Materials

Overview

On this page you will find information about the plasmids, strains, primers and DNA sequences we used. For the kits used for our experiments, the references are indicated on the protocol page.

  • • pET21 b (+) : this plasmid was used as a backbone for all constructs. Its origin of replication is derived from pBR322. This plasmid confers ampicillin resistance. This pET21 b (+) was a gift from Ambre Jousselin (addgene catalog number: 69741-3).

  • • psb1c3_double-inverse_ihfB800BFP_xylaRFP : this plasmid was used to recover the ihfB800 promoter with mTagBFP sequence by PCR. This plasmid was a gift from Manon Barthe.

  • • psb1c3_double-inverse_ihfB800RFP_xylaBFP : this plasmid was used to recover the ihfB800 promoter with mRFP1 sequence by PCR. This plasmid was a gift from Manon Barthe.

  • • pVIBI_1_mSCARLET-I : this plasmid was used to recover the mScarlet-I sequence by PCR. This plasmid was a gift from Maxence Holtz.

  • • E. coli Stellar : (F–, endA1, supE44, thi-1, recA1, relA1, gyrA96, phoA, Φ80d lacZΔ M15, Δ(lacZYA-argF) U169, Δ(mrr-hsdRMS-mcrBC), ΔmcrA, λ–) used for transformation during In-Fusion® cloning (Stellar Competent cells from Takara, ref : 636763).

  • • E. coli (DE3) tuner : (F– ompT hsdSB (rB– mB–) gal dcm lacY1(DE3)) for the expression of our sequence. The strain is a lacZY deletion mutant of BL21. Protein production is induced by IPTG, which enters into all cells uniformly in the population through the lac permease (lacY) mutation, thus allowing a homogeneous and linear response to the amount of IPTG. For this strain, the gene encoding for the T7 polymerase was integrated into the chromosome under the control of the IPTG-inducible lacUV5 promoter. This strain was a gift from Cédric Montanier.

Amplification of gBlocks

Table 1 : Set of primers used for gBlocks amplification

gBlocks name Name Sequence
ompA-DARPin

IF3_allergen-F

gccgcaagctttaatgatggtgatggtgatggtgatg

IF4_GalD2/DARPin-R

cgagctccgtcgacaaggaggtaatatacatatgaaagcc
ompA-Gal d 2

IF3_allergen-F

gccgcaagctttaatgatggtgatggtgatggtgatg

IF4_GalD2/DARPin-R

cgagctccgtcgacaaggaggtaatatacatatgaaagcc
Ara h 2

IF3_allergen-F

gccgcaagctttaatgatggtgatggtgatggtgatg

IF4_Arah2

cctgtattttcagagcatggccaagctcaccattc
Ana o 3

IF3_allergen-F

gccgcaagctttaatgatggtgatggtgatggtgatg

IF4_Anao3

cctgtattttcagagcatggcgaaatttcttttattattg
Der p 1

IF3_allergen-F

gccgcaagctttaatgatggtgatggtgatggtgatg

IF4_Derp1

cctgtattttcagagcatgaaaatcgtgctggc

Amplification of fluorophore sequences

Table 2 : Primer pairs to amplify fluorescent sequences

Fluorophore name Name Sequence
mTagBFP

mTagBFP/mRFP1 insertion with BglII site creation (F)

cgcgggatcgagatctatcacgaggcagaatttcagat

mTagBFP/mRFP1 insertion 2 (R)

tagaggatcgagatctctgaaacagtgcaaagctaaccc
mRFP1

mTagBFP/mRFP1 insertion with BglII site creation (F)

cgcgggatcgagatctatcacgaggcagaatttcagat

mTagBFP/mRFP1 insertion 2 (R)

tagaggatcgagatctctgaaacagtgcaaagctaaccc
Exclusion of mRFP1

IF2_exclusion_mRFP1 (F)

acctttagaaaccatctctaggattcctccgg

IF1_exclusion_mRFP1 (R)

gaactgtacaaataatctagaagcggccgcgaattc
mScarlet-I

IF3_mSCARLET-I (F)

ttatttgtacagttcatccataccacc

F4_mSCARLET-I (R)

atggtttctaaaggtgaagcagtg

Screening

Table 3 : Screening primers

gBlocks name Name Sequence
ompA-DARPin

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc
ompA-Gal d 2

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc
Ara h 2

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc
Ana o 3

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc
Der p 1

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc
mTagBFP

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc
mRFP1

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc

For plasmid linearization

Table 4 : Primer pairs to linearize plasmids

Construction name Name Sequence
pET21 b (+)

IF1_GalD2/DARPin-F

tgtcgacggagctcgaattcg

IF2_plasmid-R

ttaaagcttgcggccgcactcg
pET21 b (+)_OmpA without Gal d 2

IF1_allergen

gctctgaaaatacaggttttcactg

F2_plasmid-R

ttaaagcttgcggccgcactcg
pET21 b (+)_OmpA_DARPin_sfGFP

exclusion sfGFP by In-Fusion (F)

tggtgatgcagtttctgcaaaatttccgcc

exclusion sfGFP by In-Fusion (R)

gaaactgcatcaccatcaccatcacc
pET21 b (+)_OmpA_DARPin not complete_ihfB800_BFP

F1 plasmid DARPin (F)

gccgagatcgctaccgctctgaaaatacaggttttcactg

IF2 plasmid DARPin (R)

ctgcatcttgcagccatgctaggccatctggaaattg

Checking insert sequence

Table 5 : Primers used to check inserts

Sequence name Name Sequence
ompA-DARPin

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc

seq_GalD2-F1

tcttgctcagtcactcggaac

seq_GalD2-F2

catggtggactttaagctcc
ompA-Gal d 2

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc

seq_DARPin-F1

atcagggtagcgcgagaaac

seq_DARPin-F2

gcctaaccgagtgtagatgtc
Ara h 2

screening_insert-F

ggttatgctagttattgctcagc

IF1_allergen

gctctgaaaatacaggttttcactg

seq_Arah2-F1

atcgagatctcgatcccgc
Ana o 3

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc

IF1_allergen

gctctgaaaatacaggttttcactg
Der p 1

screening_insert-F

ggttatgctagttattgctcagc

screening_insert-R

ccgaaacaagcgctcatgagc

seq_Derp1-F1

ctgtgatgcacagtcgacaag

seq_Derp1-F2

ttgctcttggtatctgcgc
mRFP1

IF1_GalD2/DARPin-F

tgtcgacggagctcgaattcg

sequençage mRFP1

gacagtttcaggtagtccgggatg

screening_insert-R

ccgaaacaagcgctcatgagc
mTagBFP

IF1_GalD2/DARPin-F

tgtcgacggagctcgaattcg

sequençage mTagBFP1

gccacgaattttcacgttatag

screening_insert-R

ccgaaacaagcgctcatgagc

DARPin missing part reconstitution

Table 6 : Polymerase cycling assembly and amplification primer pairs to reconstitute the missing part of DARPin

Goal Name Sequence
Reconstruction of the missing part of DARPin by PCA

1_PCA50 (F)

ggctgcaagatgcagaggtgtgtggccccaatagtcattcgcgttgacgtccgcccc

2_PCA50 (R)

cgcgttgacgtccgccccgttggcggtcagaatgcgaacttcatcgtcttgtccggcgcg

3_PCA50 (R)

cttcatcgtcttgtccggcgcgagccgcttcgagtaatttcttgccgagatcgctacc
Amplification of the missing part of DARPin

IF3 ampli DARPin (F)

ggctgcaagatgcagaggtg

IF4 ampli DARPin (R)

ggtagcgatctcggcaagaaattac

Plasmids

pET21 b (+) :

atccggatatagttcctcctttcagcaaaaaacccctcaagacccgtttagaggccccaaggggttatgctagttattgctcagcggtggcagcagccaactcagcttcctttcgggctttgttagcagccggatctcagtggtggtggtggtggtgctcgagtgcggccgcaagcttgtcgacggagctcgaattcggatcccgacccatttgctgtccaccagtcatgctagccatatgtatatctccttcttaaagttaaacaaaattatttctagaggggaattgttatccgctcacaattcccctatagtgagtcgtattaatttcgcgggatcgagatctcgatcctctacgccggacgcatcgtggccggcatcaccggcgccacaggtgcggttgctggcgcctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcgcttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggcgccatctccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctactactgggctgcttcctaatgcaggagtcgcataagggagagcgtcgagatcccggacaccatcgaatggcgcaaaacctttcgcggtatggcatgatagcgcccggaagagagtcaattcagggtggtgaatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagaccgtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaagcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaacagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaattgtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggtagaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtcagtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcctgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtattattttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcaccagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggctggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgactggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttcccactgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgagtccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagctcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaaccagcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttgcccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtaagttagctcactcattaggcaccgggatctcgaccgatgcccttgagagccttcaacccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcacttatgactgtcttctttatcatgcaactcgtaggacaggtgccggcagcgctctgggtcattttcggcgaggaccgctttcgctggagcgcgacgatgatcggcctgtcgcttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcactggtcccgccaccaaacgtttcggcgagaagcaggccattatcgccggcatggcggccccacgggtgcgcatgatcgtgctcctgtcgttgaggacccggctaggctggcggggttgccttactggttagcagaatgaatcaccgatacgcgagcgaacgtgaagcgactgctgctgcaaaacgtctgcgacctgagcaacaacatgaatggtcttcggtttccgtgtttcgtaaagtctggaaacgcggaagtcagcgccctgcaccattatgttccggatctgcatcgcaggatgctgctggctaccctgtggaacacctacatctgtattaacgaagcgctggcattgaccctgagtgatttttctctggtcccgccgcatccataccgccagttgtttaccctcacaacgttccagtaaccgggcatgttcatcatcagtaacccgtatcgtgagcatcctctctcgtttcatcggtatcattacccccatgaacagaaatcccccttacacggaggcatcagtgaccaaacaggaaaaaaccgcccttaacatggcccgctttatcagaagccagacattaacgcttctggagaaactcaacgagctggacgcggatgaacaggcagacatctgtgaatcgcttcacgaccacgctgatgagctttaccgcagctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgcgtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatatatgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttgccattgctgcaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgaaattgtaaacgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcattttttaaccaataggccgaaatcggcaaaatcccttataaatcaaaagaatagaccgagatagggttgagtgttgttccagtttggaacaagagtccactattaaagaacgtggactccaacgtcaaagggcgaaaaaccgtctatcagggcgatggcccactacgtgaaccatcaccctaatcaagttttttggggtcgaggtgccgtaaagcactaaatcggaaccctaaagggagcccccgatttagagcttgacggggaaagccggcgaacgtggcgagaaaggaagggaagaaagcgaaaggagcgggcgctagggcgctggcaagtgtagcggtcacgctgcgcgtaaccaccacacccgccgcgcttaatgcgccgctacagggcgcgtcccattcgcca

Table 7 : Features of pET21 b (+)

Feature name End Start
T7 terminator

1

129
MCS

230

307
T7 promoter

308

326
pBR322_origin

3210

3829
Ampicillin resistance gene

3984

4844

psb1c3_double-inverse_ihfB800BFP_xylaRFP

cttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgcccgatcaactcgagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggcagaatttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagattattaattcagtttatgacccagcttgctaggcaagtcgcaatagcgcgcgactgcaacctcgtgctgctccacgtaagtctcattattcgcttctttaatgcgttccagacggtagtcgacatagtaaacacccggcatcttcagatttttcgccggtttcttgctgcggtacgtcgtcttgatatttgcgatcaaatggctgccaccaaccagcttcagtgccatgtcgttacggccctccaggccaccatcggccggatacagggtttccgtaaacgcctcccaacccagggttttcttctgcatcaccggaccattgctcgtaaagttgacgccacgaattttcacgttatagatcagacagccatcttgcagggaggtatcttgggtggcggtcaacacaccaccatcttcatacgtggtgacacgctcccaggtgaaaccttccggaaagctctgtttgaagaaatccggaatgccctgagtatggttaatgaacgttttggaaccgtacaggaagctggtcgccagaatgtcaaaagcgaacggcagtggaccgccctccacaactttgatacgcatggtttgggtgccttcgtacggcttaccctcaccctcagacgtgcacttaaagtggtggttgtcaacggtgccttccatgtacagcttcatgtgcatgttctctttgatcagttcgctcatctctaggattcctccggttccttaaaggcaaattaagcggctgctgttgctgcattgattaagggcggccgtagccgcccttagtgcttgattacaggacgaaacctgcaatctgtcaagtaaactcaacaaacttcggaataaaaatcccgaagagtcagagaattactcgcctttagctgctttgaaagcttcagccattgcgttgttggagaagtttgcatcttcctgtttgttaacagttgcgattgcatctttctcgtcagcttcgtctttcgcacgaacagacaggctgattgcgcggtttttacgatcaacgccggtgaatttagcttcaacttcgtcgccaacgctcagaaccagggtagcgtcttcaacgcggtcacgggatgcttcagaagcacgcaggtaaccttcaacgccgtcagccagttctacggttgcgcctttagcgtcaactgcagttactttaccggttacgatagcgcctttcttgttcagagcaacccagttgttgaacggatcttctgcgagctgtttaacgcccagggagatacgttcacgttctgcgtcaacctgcagaacaactgcagcgatttcgtcgccttttttgtattcacgaactgcttcttcgcctgcaacgttccaggagatgtcagacaggtgaaccaggccgtcgatgccgccgtccaagccgatgaagataccgaagtcagtgatagacttgattttaccttcaacacggtcgcccttgttgtgggtttccgcgaactgctgccacgggttagctttgcactgtttcagaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatactagaatttttagcaactaaacaggggaaaacaattacagatttttatctttcgattacgatttttggtttatttcttgatttatgaccgagatcttacttttgttgcgcaattgtacttattgcatttttctcttcgaggaattacccagtttcatcattccattttattttgcgagcgagcgcacacttgtgaattatctcaatagcagtgtgaaataacataattgagcaactgaaagggagtgcccaatattacgacatcatccatcacccgcggcattacctgattatggagttcaatactagagatggcttcctccgaagacgttatcaaagagttcatgcgtttcaaagttcgtatggaaggttccgttaacggtcacgagttcgaaatcgaaggtgaaggtgaaggtcgtccgtacgaaggtacccagaccgctaaactgaaagttaccaaaggtggtccgctgccgttcgcttgggacatcctgtccccgcagttccagtacggttccaaagcttacgttaaacacccggctgacatcccggactacctgaaactgtccttcccggaaggtttcaaatgggaacgtgttatgaacttcgaagacggtggtgttgttaccgttacccaggactcctccctgcaagacggtgagttcatctacaaagttaaactgcgtggtaccaacttcccgtccgacggtccggttatgcagaaaaaaaccatgggttgggaagcttccaccgaacgtatgtacccggaagacggtgctctgaaaggtgaaatcaaaatgcgtctgaaactgaaagacggtggtcactacgacgctgaagttaaaaccacctacatggctaaaaaaccggttcagctgccgggtgcttacaaaaccgacatcaaactggacatcacctcccacaacgaagactacaccatcgttgaacagtacgaacgtgctgaaggtcgtcactccaccggtgcttaataacgctgatagtgctagtgtagatcgctactagtagcggccgctgcagtccggcaaaaaagggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgttatgcaggcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccacaggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagctcgaggcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggatctatcaacaggagtccaagcgagctcgatatcaaattacgccccgccctgccactcatcgcagtactgttgtaattcattaagcattctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacgaaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttag


Table 8 : Features of psb1c3_double-inverse_ihfB800BFP_xylaRFP

Feature name End Start
Bacterial terminator

179

222
mTagBFP

246

950
ihfB800 promoter

957

1755
Ori

3762

3174
Chloramphenicol resistance gene

4063

4719

psb1c3_double-inverse_ihfB800RFP_xylaBFP :

cgccccgccctgccactcatcgcagtactgttgtaattcattaagcattctgccgacatggaagccatcacaaacggcatgatgaacctgaatcgccagcggcatcagcaccttgtcgccttgcgtataatatttgcccatggtgaaaacgggggcgaagaagttgtccatattggccacgtttaaatcaaaactggtgaaactcacccagggattggctgagacgaaaaacatattctcaataaaccctttagggaaataggccaggttttcaccgtaacacgccacatcttgcgaatatatgtgtagaaactgccggaaatcgtcgtggtattcactccagagcgatgaaaacgtttcagtttgctcatggaaaacggtgtaacaagggtgaacactatcccatatcaccagctcaccgtctttcattgccatacgaaattccggatgagcattcatcaggcgggcaagaatgtgaataaaggccggataaaacttgtgcttatttttctttacggtctttaaaaaggccgtaatatccagctgaacggtctggttataggtacattgagcaactgactgaaatgcctcaaaatgttctttacgatgccattgggatatatcaacggtggtatatccagtgatttttttctccattttagcttccttagctcctgaaaatctcgataactcaaaaaatacgcccggtagtgatcttatttcattatggtgaaagttggaacctcttacgtgcccgatcaactcgagtgccacctgacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgaggcagaatttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagagcgatctacactagcactatcagcgttattaagcaccggtggagtgacgaccttcagcacgttcgtactgttcaacgatggtgtagtcttcgttgtgggaggtgatgtccagtttgatgtcggttttgtaagcacccggcagctgaaccggttttttagccatgtaggtggttttaacttcagcgtcgtagtgaccaccgtctttcagtttcagacgcattttgatttcacctttcagagcaccgtcttccgggtacatacgttcggtggaagcttcccaacccatggtttttttctgcataaccggaccgtcggacgggaagttggtaccacgcagtttaactttgtagatgaactcaccgtcttgcagggaggagtcctgggtaacggtaacaacaccaccgtcttcgaagttcataacacgttcccatttgaaaccttccgggaaggacagtttcaggtagtccgggatgtcagccgggtgtttaacgtaagctttggaaccgtactggaactgcggggacaggatgtcccaagcgaacggcagcggaccacctttggtaactttcagtttagcggtctgggtaccttcgtacggacgaccttcaccttcaccttcgatttcgaactcgtgaccgttaacggaaccttccatacgaactttgaaacgcatgaactctttgataacgtcttcggaggaagccatctctaggattcctccggttccttaaaggcaaattaagcggctgctgttgctgcattgattaagggcggccgtagccgcccttagtgcttgattacaggacgaaacctgcaatctgtcaagtaaactcaacaaacttcggaataaaaatcccgaagagtcagagaattactcgcctttagctgctttgaaagcttcagccattgcgttgttggagaagtttgcatcttcctgtttgttaacagttgcgattgcatctttctcgtcagcttcgtctttcgcacgaacagacaggctgattgcgcggtttttacgatcaacgccggtgaatttagcttcaacttcgtcgccaacgctcagaaccagggtagcgtcttcaacgcggtcacgggatgcttcagaagcacgcaggtaaccttcaacgccgtcagccagttctacggttgcgcctttagcgtcaactgcagttactttaccggttacgatagcgcctttcttgttcagagcaacccagttgttgaacggatcttctgcgagctgtttaacgcccagggagatacgttcacgttctgcgtcaacctgcagaacaactgcagcgatttcgtcgccttttttgtattcacgaactgcttcttcgcctgcaacgttccaggagatgtcagacaggtgaaccaggccgtcgatgccgccgtccaagccgatgaagataccgaagtcagtgatagacttgattttaccttcaacacggtcgcccttgttgtgggtttccgcgaactgctgccacgggttagctttgcactgtttcagaccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttatactagaatttttagcaactaaacaggggaaaacaattacagatttttatctttcgattacgatttttggtttatttcttgatttatgaccgagatcttacttttgttgcgcaattgtacttattgcatttttctcttcgaggaattacccagtttcatcattccattttattttgcgagcgagcgcacacttgtgaattatctcaatagcagtgtgaaataacataattgagcaactgaaagggagtgcccaatattacgacatcatccatcacccgcggcattacctgattatggagttcaatactagagatgagcgaactgatcaaagagaacatgcacatgaagctgtacatggaaggcaccgttgacaaccaccactttaagtgcacgtctgagggtgagggtaagccgtacgaaggcacccaaaccatgcgtatcaaagttgtggagggcggtccactgccgttcgcttttgacattctggcgaccagcttcctgtacggttccaaaacgttcattaaccatactcagggcattccggatttcttcaaacagagctttccggaaggtttcacctgggagcgtgtcaccacgtatgaagatggtggtgtgttgaccgccacccaagatacctccctgcaagatggctgtctgatctataacgtgaaaattcgtggcgtcaactttacgagcaatggtccggtgatgcagaagaaaaccctgggttgggaggcgtttacggaaaccctgtatccggccgatggtggcctggagggccgtaacgacatggcactgaagctggttggtggcagccatttgatcgcaaatatcaagacgacgtaccgcagcaagaaaccggcgaaaaatctgaagatgccgggtgtttactatgtcgactaccgtctggaacgcattaaagaagcgaataatgagacttacgtggagcagcacgaggttgcagtcgcgcgctattgcgacttgcctagcaagctgggtcataaactgaattaataatactagtagcggccgctgcagtccggcaaaaaagggcaaggtgtcaccaccctgccctttttctttaaaaccgaaaagattacttcgcgttatgcaggcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccacaggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagctcgaggcttggattctcaccaataaaaaacgcccggcggcaaccgagcgttctgaacaaatccagatggagttctgaggtcattactggatctatcaacaggagtccaagcgagctcgatatcaaatta


Table 9 : Features of psb1c3_double-inverse_ihfB800RFP_xylaBFP

Feature name End Start
CmR gene

1

657
Bacterial terminator

841

884
mRFP1

908

1613
ihfB800 promoter

1620

2418
Ori

4424

3836

DARPin and Allergen gBlocks

ompA-DARPin sequence with sfGFP and HIS-tag :

ttaatgatggtgatggtgatggtgatgtttgtataattcatccatgccatgggtaatacccgccgctgtaacaaattccagcaggaccatgtgatcccgcttctcatttggatcttttgaaagtacactttgtgtcgacaaataatgattgtccggcagcagcacggggccatcgccaataggggtattctgctgataatgatcggccagttggacagagccatcttcaacgttatgacggattttgaaatttgctttaatgccatttttttgtttgtctgcCgtgatatacacgttatgactgttaaagttatattctagtttatgcccaagtatgttaccgtcttctttgaagtcaatacctttcagttcaatgcggttaactaacgtatcgccttcaaacttgacttcagcgcgggttttgtaggttccgtcatccttaaaggagatggtacgttcctgcacgtaaccctccggcatcgcgcttttgaaaaaatcgtgctgtttcatgtgatcagggtagcgcgagaaacactgtaccccataagtcagcgtcgtgaccaacgtcggccacggcactggcagtttaccagtggtgcaaataaacttgagggtgagttttccattcgtggcgtcgccctcaccttcaccacgtacgctaaacttgtgcccgttcacatctccgtccagctcaaccaggatcggaaccacgcctgtgaacaactcttcgcctttgctactgctaatgcccagtttctgcaaaatttccgccagatcctcgttcccattgtcgatgctgatgtcgaacgcagttttaccaaacttatcttgggcgttaacgtctgctccatgtttgagcagcacctcgacaatttcaaggtgatcagcccaggcagccaagtgcagcggggtgcgaccggtattgcccgtcgcgtttacatctgcgccatttttcagtaaaacttccacaatttccagatggcctagcatggctgcaagatgcagaggtgtgtggccccaatagtcattcgcgttgacgtccgccccgttggcggtcagaatgcgaacttcatcgtcttgtccggcgcgagccgcttcgagtaatttcttgccgagatcgctaccgctctgaaaatacaggttttcactgctaatgccacgcgtcgcaatttcgggggtgatcgcatactcgacacccccggcgaaaactggactcacccccgtatcatgatttttgccatatacgttgctcttggtatctgcgcgccacaccatgccgcctaaccgagtgtagatgtccaggtcatcagtaatagggtagccaagtttcgctgtcaactgaacgccctgtgctttataagccccgttctctacggaacctttgtacggcatgcgtcccagccaatcatagcccatttcaaatcccacgtacgggttgacctgataaccaccaaaggcaccagcacccagttggttttcgtgggtcgggccattattattctgatcaattttcgcgttgctactgcaccctgccagcagcgtgctacctaaaatcaccgcgcccaaaacaagtttggtggctttcatatgtatattacctcct


Table 10 : Features of ompA-DARPin sequence with sfGFP and HIS-tag

Feature name End Start
Stop codon

1

3
HIS-tag

4

27
sfGFP

28

738
GISS linker

739

750
DARPin e_79

751

1125
TEV clivage site

1126

1146
GISS linker

1147

1158
ompA

1159

1587
RBS

1595

1603

ompA-Gal d 2 sequence and HIS-tag :

ttaatgatggtgatggtgatggtgatgactgctaatgcccggggaaacacagcggccaaagaagagaacggcgttggttgcgatgtgtttgatacagaacagaaacggatggtcagcgcgaaattcttccgagacgcttgccgcatccactccagcttccgctgaccctaccacttcacggcctgcttcattaatttctgcatgcgccgcatggaccgcttgagatatcttcaggctctctgctgaggagatgccagacagattggctgaggagctaaacacgtccgtaatgcccatagccattaagacagacgtcaggttatatttttcttccattttcatgcgaggtaagtacactttaatcttgcgttcttccataacattcgaactggtccattccgtcagtttttcaaagttaataatactctcaagctgctccagccctgagacttcatcaggcaacagcaccaacatgctcattgtcccactggcaaatggcagctccagaatcttcattttctccgaagccatcgatgccactctaaataaaccaatctggtacatcatctgcactggtttgctttcttgctcagtcactcggaacggcatcgcctgtgtgtcttcatccttaaacgctttctcccacagacctttgaagacaatggcattcaccagaaccatcgcagtctgagaatccacggagctcggttgcaggacattacggataataccgttggtctgactttctacccaggaattgatgagctcccgggcttgatctgccgcggtttgaaagttgatcggttccaagcctccgcgatacagttctttcacgcactgcaagtattccggcaggatgggataacgctcttccgcatataaacgactggccaggctgaacgaataaacatcgttcggtttggtgatttggttgaggatgtcccgaaggctagagtgaacatttaccgaagtgccgcactgagcttcaatactgtcgccgaaacctggaagtttatcaaagcgaacaactttatttatctgcgtacgggtgctatctttagcacccaggtataccatggctagcgctgacatgatggcaatggggcaatagaaaatgttctcgttggcatggtggactttaagctccttgaatacatcaaaacaaaattccatgcttgccgcaccaatggagcccatgctctgaaaatacaggttttcactgctaatgccacgcgtcgcaatttcgggggtgatcgcatactcgacacccccggcgaaaactggactcacccccgtatcatgatttttgccatatacgttgctcttggtatctgcgcgccacaccatgccgcctaaccgagtgtagatgtccaggtcatcagtaatagggtagccaagtttcgctgtcaactgaacgccctgtgctttataagccccgttctctacggaacctttgtacggcatgcgtcccagccaatcatagcccatttcaaatcccacgtacgggttgacctgataaccaccaaaggcaccagcacccagttggttttcgtgggtcgggccattattattctgatcaattttcgcgttgctactgcaccctgccagcagcgtgctacctaaaatcaccgcgcccaaaacaagtttggtggctttcatatgtatattacctcct


Table 11 : Features of ompA-Gal d 2 sequence and HIS-tag

Feature name End Start
Stop codon

1

3
HIS-tag

4

27
GISS linker

28

39
Gal d 2

40

1197
TEV clivage site

1198

1218
GISS linker

1219

1230
ompA

1231

1659
RBS

1667

1675

Ara h 2 sequence with HIS-tag :

ttaatgatggtgatggtgatggtgatgactgctaatgccgtatcgatcgcggccaccggattcaacttccaggtcgcaacgctgtggtgcgcgaaggccgcactgttgaggcaagttgcgtagttcgcgtttgaactgttgctcctgttgacgcccctgcaagcgatcgctctgattctccataatctgttgtaatgcttcacacatacagcgctgattgttttcaaattcgttcagttcattgcaacagcgctcttgatgctgagacgaaccagcgccacgccgatcatatggactcgggctgtacggatcacggcgatccgggtcctgactaggtgaatacggatcctgcgacgggctgtacggatcccgaccatatgaatcttcgtcacgttggattttctgcattaaatgttgttcacagggcctcaggttcgcacgctccagctgggactggcatctacggtctccttggagttcccactgctgacgcgcagacgcgtgcgcagccagcagaaaaagggccagggccaccagaatggtgagcttggccat


Table 12 : Features of Ara h 2 sequence with HIS-tag

Feature name End Start
Stop codon

1

3
HIS-tag

4

27
GISS linker

28

39
Ara h 2

40

555

Ana o 3 sequence with HIS-tag :

atggcgaaatttcttttattattgagcgctttcgcagtactgctgctggttgcaaatgcgtcgatttatcgcgccattgtggaggtagaagaagactccggtcgtgagcagtcatgtcagcgtcaattcgaagagcagcagcggtttcgtaattgtcagcgctatgttaaacaagaagtccagcgtggcgggcggtacaaccaacgccaggagagtctaagagaatgctgccaggaactgcaagaggtcgatcgccgttgtcgctgtcagaacctggaacaaatggtgcgccagctccagcagcaagaacagatcaagggtgaggaagtgcgagaattgtacgaaaccgcctctgaactgccacgtatttgctccatcagcccgtcgcaaggctgccagtttcagagtagctatggcattagcagtcatcaccatcaccatcaccatcattaa


Table 13 : Features of Ana o 3 sequence with HIS-tag

Feature name End Start
Stop codon

1

3
HIS-tag

4

27
GISS linker

28

39
Ana o 3

40

453

Der p 1 sequence with HIS-tag :

ttaatgatggtgatggtgatggtgatgactgctaatgcctaatatcaccacataaggatactcttcaatcatcattaagtcaatattggctgcaaagtaaccatagccgttatcaccccaattcgtatcccatgagttacgaacgatccaataatccaccccctgagcattactatagccaactatattaaccgcatggtagttcggctgataaccgttatcacgctggataatggtacggccatcgtaatgcctaaatgcgtccaggtctttaataccaataatgacagcgatcgcagagtgggtttgtgccagcgcttcacggatcttattgacgtttggcggataaatctggcaatagttactaatgccgaagcgctgcgcgttcggacgccgacaggattgttcgcgtgctacatagcgatagtagctttcctgaactacgccattgtgttggatgtattcaatgccgcgtggaatggtatccccatggcatccatgctgtgatgcacagtcgacaagttcttgttccgctaagtcaagactctggtttctgtaggccaggtacgcgctttcagtcgcagccacgcccgaaaaggcccagcagcttccgcaacccccttgcatgcggatgggagttactgtacgcatctggcgcaagtcaatttcagccggagcgttaccgttgatgctacaggcattcgtctcggcattcaaatcaaactgtgttttcagatgctcaaaggcttctgctgacatcagaaatcggttcttaaattcatccagagagagatcggacaggtgattgattgcgccaccattcgactgcacgtatttgacggattccaggaagtttttgcgtgcggcttcctcatcctcgaacgtggcgtagctcttattgaacgcttttttatactcttcaaaggttttgatgctactcggccgcgcataaaccgcagacagcgcaaggagcgacgcaatggccagcacgattttcat

Table 14 : Features of Der p 1 sequence with HIS-tag

Feature name End Start
Stop codon

1

3
HIS-tag

4

27
GISS linker

28

39
Der p 1

40

999

Fluorophore sequences

Sequence with ihfB800 promoter, mRFP1 gene and bacterial terminator :

atttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagagcgatctacactagcactatcagcgttattaagcaccggtggagtgacgaccttcagcacgttcgtactgttcaacgatggtgtagtcttcgttgtgggaggtgatgtccagtttgatgtcggttttgtaagcacccggcagctgaaccggttttttagccatgtaggtggttttaacttcagcgtcgtagtgaccaccgtctttcagtttcagacgcattttgatttcacctttcagagcaccgtcttccgggtacatacgttcggtggaagcttcccaacccatggtttttttctgcataaccggaccgtcggacgggaagttggtaccacgcagtttaactttgtagatgaactcaccgtcttgcagggaggagtcctgggtaacggtaacaacaccaccgtcttcgaagttcataacacgttcccatttgaaaccttccgggaaggacagtttcaggtagtccgggatgtcagccgggtgtttaacgtaagctttggaaccgtactggaactgcggggacaggatgtcccaagcgaacggcagcggaccacctttggtaactttcagtttagcggtctgggtaccttcgtacggacgaccttcaccttcaccttcgatttcgaactcgtgaccgttaacggaaccttccatacgaactttgaaacgcatgaactctttgataacgtcttcggaggaagccatctctaggattcctccggttccttaaaggcaaattaagcggctgctgttgctgcattgattaagggcggccgtagccgcccttagtgcttgattacaggacgaaacctgcaatctgtcaagtaaactcaacaaacttcggaataaaaatcccgaagagtcagagaattactcgcctttagctgctttgaaagcttcagccattgcgttgttggagaagtttgcatcttcctgtttgttaacagttgcgattgcatctttctcgtcagcttcgtctttcgcacgaacagacaggctgattgcgcggtttttacgatcaacgccggtgaatttagcttcaacttcgtcgccaacgctcagaaccagggtagcgtcttcaacgcggtcacgggatgcttcagaagcacgcaggtaaccttcaacgccgtcagccagttctacggttgcgcctttagcgtcaactgcagttactttaccggttacgatagcgcctttcttgttcagagcaacccagttgttgaacggatcttctgcgagctgtttaacgcccagggagatacgttcacgttctgcgtcaacctgcagaacaactgcagcgatttcgtcgccttttttgtattcacgaactgcttcttcgcctgcaacgttccaggagatgtcagacaggtgaaccaggccgtcgatgccgccgtccaagccgatgaagataccgaagtcagtgatagacttgattttaccttcaacacggtcgcccttgttgtgggtttccgcgaactgctgccacgggttagctttgcactgtttcaga


Table 15 : Features of sequence with ihfB800 promoter, mRFP1 gene and bacterial terminator

Feature name End Start
Bacterial terminator

1

1578
mRFP1

68

773
ihfB800

780

1578

Sequence with ihfB800 promoter, mTagBFP gene and bacterial terminator :


atttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagattattaattcagtttatgacccagcttgctaggcaagtcgcaatagcgcgcgactgcaacctcgtgctgctccacgtaagtctcattattcgcttctttaatgcgttccagacggtagtcgacatagtaaacacccggcatcttcagatttttcgccggtttcttgctgcggtacgtcgtcttgatatttgcgatcaaatggctgccaccaaccagcttcagtgccatgtcgttacggccctccaggccaccatcggccggatacagggtttccgtaaacgcctcccaacccagggttttcttctgcatcaccggaccattgctcgtaaagttgacgccacgaattttcacgttatagatcagacagccatcttgcagggaggtatcttgggtggcggtcaacacaccaccatcttcatacgtggtgacacgctcccaggtgaaaccttccggaaagctctgtttgaagaaatccggaatgccctgagtatggttaatgaacgttttggaaccgtacaggaagctggtcgccagaatgtcaaaagcgaacggcagtggaccgccctccacaactttgatacgcatggtttgggtgccttcgtacggcttaccctcaccctcagacgtgcacttaaagtggtggttgtcaacggtgccttccatgtacagcttcatgtgcatgttctctttgatcagttcgctcatctctaggattcctccggttccttaaaggcaaattaagcggctgctgttgctgcattgattaagggcggccgtagccgcccttagtgcttgattacaggacgaaacctgcaatctgtcaagtaaactcaacaaacttcggaataaaaatcccgaagagtcagagaattactcgcctttagctgctttgaaagcttcagccattgcgttgttggagaagtttgcatcttcctgtttgttaacagttgcgattgcatctttctcgtcagcttcgtctttcgcacgaacagacaggctgattgcgcggtttttacgatcaacgccggtgaatttagcttcaacttcgtcgccaacgctcagaaccagggtagcgtcttcaacgcggtcacgggatgcttcagaagcacgcaggtaaccttcaacgccgtcagccagttctacggttgcgcctttagcgtcaactgcagttactttaccggttacgatagcgcctttcttgttcagagcaacccagttgttgaacggatcttctgcgagctgtttaacgcccagggagatacgttcacgttctgcgtcaacctgcagaacaactgcagcgatttcgtcgccttttttgtattcacgaactgcttcttcgcctgcaacgttccaggagatgtcagacaggtgaaccaggccgtcgatgccgccgtccaagccgatgaagataccgaagtcagtgatagacttgattttaccttcaacacggtcgcccttgttgtgggtttccgcgaactgctgccacgggttagctttgcactgtttcaga


Table 16 : Features of sequence with ihfB800 promoter, mTagBFP gene and bacterial terminator

Feature name End Start
Bacterial terminator

1

44
mTagBFP

68

772
ihfB800

779

1577

Sequence with ihfB800 promoter, mScarlet-I gene and bacterial terminator :

atttcagataaaaaaaatccttagctttcgctaaggatgatttctggaattcgcggccgcttctagattatttgtacagttcatccataccaccggtagagtgacggccttcagaacgttcgtattgttcaacaacggtgtaatcttcgttgtggctggtaatatccagtttacgatcaacgttgtatgcacctggcatttgaactggttttttcgctttgtaggtggttttgaaatctgccaggtaacgaccgccatctttcagacgcagtgccattttgatatcacctttcagaacgccatcttctgggtacagacgttcggtagatgcttcccaacccatggtttttttctgcataactggaccatctggtggaaagttagtaccacgcagtttaactttgtagatcagagtaccatcttccagagaggtatcttgggtaaccgtaactgcaccaccatcttcaaaattcataacacgttcccatttgaaaccttccggaaaagattgtttgtagtagtctggaatatctgctgggtgtttgataaatgcacgagaaccgtacataaattgtggagacagaatatcccaagaaaatggcagtggaccacctttggtaactttcagttttgcggtttgagtaccttcgtatggacgaccttcgccttcaccttcaatttcaaattcgtggccgttcatagaaccttccatgtgaactttgaagcgcataaattctttgatcactgcttcacctttagaaaccatctctaggattcctccggttccttaaaggcaaattaagcggctgctgttgctgcattgattaagggcggccgtagccgcccttagtgcttgattacaggacgaaacctgcaatctgtcaagtaaactcaacaaacttcggaataaaaatcccgaagagtcagagaattactcgcctttagctgctttgaaagcttcagccattgcgttgttggagaagtttgcatcttcctgtttgttaacagttgcgattgcatctttctcgtcagcttcgtctttcgcacgaacagacaggctgattgcgcggtttttacgatcaacgccggtgaatttagcttcaacttcgtcgccaacgctcagaaccagggtagcgtcttcaacgcggtcacgggatgcttcagaagcacgcaggtaaccttcaacgccgtcagccagttctacggttgcgcctttagcgtcaactgcagttactttaccggttacgatagcgcctttcttgttcagagcaacccagttgttgaacggatcttctgcgagctgtttaacgcccagggagatacgttcacgttctgcgtcaacctgcagaacaactgcagcgatttcgtcgccttttttgtattcacgaactgcttcttcgcctgcaacgttccaggagatgtcagacaggtgaaccaggccgtcgatgccgccgtccaagccgatgaagataccgaagtcagtgatagacttgattttaccttcaacacggtcgcccttgttgtgggtttccgcgaactgctgccacgggttagctttgcactgtttcaga

Table 17 : Features of sequence with ihfB800 promoter, mScarlet-I gene and bacterial terminator

Feature name End Start
Bacterial terminator

1

44
mScarlet-I

68

766
ihfB800

773

1571


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