This year, we build up a ternary microbial symbiosis system. Three different chassis are used: Escherichia coli, Synechococcus elongatus, and Azotobacter caulinodans. We created a series of parts to engineer these three chassis. We have edited the registry page of our parts carefully. If you want to know more about our parts, please feel free to contact us at igemshanghaitech@163.com.
| Name | Short description | Long description |
|---|---|---|
| BBa_K4115000 | sfGFP | Superfolder green fluorescence protein. (We found its sequence is different from the previous ones in registry.) |
| BBa_K4115001 | sfGFP with LVA | Superfolder GFP with an LVA degradation tag on its C terminal. |
| BBa_K4115003 | PcstA | The promoter of E.coli DH5-alpha cstA gene. Its activity will be upregulated under carbon starvation conditions. |
| BBa_K4115004 | PcstA_Mutant1 | This part is a mutant of PcstA. It is more sensitive to carbon starvation compared with the original ones. |
| BBa_K4115027 | PcstA_Mutant2 | A mutant of PcstA. It is more sensitive to carbon starvation compared with the original ones. |
| BBa_K4115049 | PcstA-delta | A PcstA variant fused with a phi R73 delta activator binding site. |
| BBa_K4115018 | PyciG | A carbon starvation promoter controlled by RpoS. The native RBS of PyciG is included in this part. |
| BBa_K4115016 | PcsiE | A carbon starvation promoter controlled by RpoS and CRP. |
| BBa_K4115017 | SacC | a β-fructofuranosidase from Mannheimia succiniciproducens |
| BBa_K4115013 | TagRFP | a monomeric red (orange) fluorescent protein (We found its sequence is different from the previous ones in registry.) |
| BBa_K4115019 | glnAp2 | A part of E. coli DH5-alpha glnA gene promoter. Its activity will be upregulated by NtrC under nitrogen starvation. |
| BBa_K4115020 | relAp4 | A part of relA gene promoter. Its activity will be upregulated by NtrC under nitrogen starvation. |
| BBa_K4115009 | NS3-2 | Neutral site 3(NS3) is commonly used in homologous recombination for S.elongatus, where NS3-1 acts as an upstream homology arm. |
| BBa_K4115010 | NSI-US | An upstream homology arm from Synechococcus 2973. |
| BBa_K4115011 | NSI-DS | An downstream homology arm from Synechococcus 2973. |
| BBa_K4115005 | sps sRNA | An artificially synthetic small RNA(sRNA) that can bind the mRNA of sps. |
| BBa_K4115015 | glgA sRNA | An artificially synthetic small RNA(sRNA) that can bind the mRNA of glgA. |
| BBa_K4115012 | lac UV5 promoter | A variant of lac promoter, which does not depend on the activation of CRP. The lac operator is included in this part. |
| BBa_K4115006 | lacI | The lacI promoter, lacI CDS and a part of pET backbone containing the terminator of lacI are included in this part. |
| BBa_K4115007 | nifA_upstream | This is a stable sequence located upstream of the nifA sequence in the genome of Azorhizobium caulinodans ORS571. It is used as a one-sided homology arm to help recombine knockout of nifA on genome. |
| BBa_K4115008 | nifA_downstream | This is a stable sequence located downstream of the nifA sequence in the genome of Azorhizobium caulinodans ORS571. It is used as a one-sided homology arm to help recombine knockout of nifA on the genome. |
| BBa_K4115002 | nifA | The coding sequence of the nifA protein in the nitrogen-fixing gene cluster. It regulates nitrogen fixation-related gene expressions. |
| BBa_K4115014 | PnifH | The promoter of the nifH on the nitrogen-fixation gene cluster, which is regulated by nifA. |
| Name | Short description | Long description |
|---|---|---|
| BBa_K4115022 | J23101-B0034-sfGFP-B0015 | sfGFP is constitutively expressed at a moderately strong level. |
| BBa_K4115023 | J23101-B0034-sfGFP(with LVA)-B0015 | sfGFP is constitutively expressed at a moderately strong level. |
| BBa_K4115024 | PcstA(long version)-B0034-sfGFP(with LVA)-B0015 | A carbon starvation reporter based on PcstA (BBa_K4115003). |
| BBa_K4115030 | PcstA(long version)-B0034-sfGFP-B0015 | A carbon starvation reporter based on PcstA (BBa_K4115003). sfGFP without LVA is used in this part. |
| BBa_K4115025 | PcstA-B0034-sfGFP(with LVA)-B0015 | A carbon starvation reporter based on PcstA (BBa_K118011). |
| BBa_K4115028 | PcstA_Mutant1-B0034-sfGFP(with LVA)-B0015 | A carbon starvation reporter based on PcstA_Mutant1. |
| BBa_K4115059 | PcstA_Mutant2-B0034-sfGFP (with LVA)-B0015 | A carbon starvation reporter based on PcstA_Mutant2. |
| BBa_K4115031 | PcsiE-B0034-sfGFP-B0015 | A carbon starvation reporter based on PcsiE. sfGFP without LVA is used in this part. |
| BBa_K4115029 | PcsiE-B0034-sfGFP(with LVA)-B0015 | A carbon starvation reporter based on PcsiE. |
| BBa_K4115032 | PyciG-sfGFP-B0015 | A carbon starvation reporter based on PyciG. sfGFP without LVA is used in this part. |
| BBa_K4115033 | PyciG-sfGFP(with LVA)-B0015 | An LVA degradation tag is added to sfGFP reporter, which causes a loss of starvation response function. |
| BBa_K4115034 | glnAp2-B0034-sfGFP-B0015 | This part is designed to be a nitrogen reporter based on glnAp2. |
| BBa_K4115035 | relAp4-B0034-sfGFP-B0015 | This part is designed to be a nitrogen reporter based on relAp4. |
| BBa_K4115036 | J23101-B0034-SacC-B0015 | SacC is constitutively expressed at a moderately strong level. |
| BBa_K4115037 | J23107-B0034-SacC-B0015 | SacC is constitutively expressed at a middle level. |
| BBa_K4115038 | J23109-B0034-SacC-B0015 | SacC is constitutively expressed at a low level. |
| BBa_K4115039 | J23101-B0034-luxR-B0015-Lux pR-B0034-sfGFP-B0015 | This part can be used as a signal receiver in lux quorum sensing system. |
| BBa_K4115040 | J23101-B0034-LuxI-B0015 | This part can be used as a signal sender in lux quorum sensing system. |
| BBa_K4115041 | nifA_upstream-BleoR-nifA_downstream | This part is used to knock out the nifA. BleoR is for selection. |
| BBa_K4115042 | PnifH-sfGFP-B0015 | This part is used to test if PnifH can be regulated by nifA. sfGFP is used as a reporter. |
| BBa_K4115043 | lac UV5 promoter-nifA-PnifH-sfGFP-B0015 | This part is used to test if PnifH can be regulated by nifA. The expression of nifA can be controlled by IPTG concentration. sfGFP is used as a reporter. |
| BBa_K4115044 | J23101-B0034-tagRFP-B0015 | TagRFP is constitutively expressed at a moderately strong level. |
| BBa_K4115045 | NS3-2-lac UV5 promoter-cscB-lacI-KanR-NS3-1 | This part is used to insert a cscB expression box into the genome of S.elongatus. |
| BBa_K4115046 | NS3-2-KanR-J23101-B0034-LuxR-B0015-Lux pR-B0034-sfGFP-B0015-NS3-1 | This part is to insert the signal receiver of lux quorum sensing system into the genome of S.elongatus. |
| BBa_K4115021 | NSI-US-lacI-lac UV5 promoter-T7 promoter-glgA sRNA-MicC-Trbcl-T7 RNA polymerase-RBS-lac promoter-cmR-T7 promoter-RBS-HFQ-Trbcl-NSI-DS | This part is used to insert a glgA gene silence box into the genome of S.elongatus. The glgA gene silence can increase the sucrose output of cscB-positive S.elongatus. |
| BBa_K4115047 | pF-B0034-sfGFP-B0015 | This part is used as the negative control when checking the function of delta activator. |
| BBa_K4115048 | J23101-delta activator-B0015-pF-B0034-sfGFP-B0015 | This part is used the experimental group when checking the function of delta activator. pF is expected to be activated by the constitutively expressed delta activator. |
| BBa_K4115057 | Amplifier based on PcstA | The delta activator is expressed by PcstA, which is a carbon starvation promoter. Then the activator increases the activity of pF to form an amplifier. |
| BBa_K4115060 | Amplifier based on PcstA_Mutant1 | The delta activator is expressed by PcstA_Mutant1, which is a carbon starvation promoter. Then the activator increases the activity of pF to form an amplifier. |
| BBa_K4115058 | Positive feedback based on PcstA_delta | The delta activator is expressed by PcstA_delta, which has a higher activity. Then the activator increases the activity of PcstA_delta to form a positive feedback. |