Team:HFLS H2Z Hangzhou/sub/parts

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Team Members

Jianan Li
Qingrui Sun
Yiming Rong
Jiayue Guo
Zhiyuan Lu
Meiqi Yuan
Zhengyao Lin
Caiyi Feng
Shuyun Zhang
Tenghao Huang
Yining Huang
Yanyue Zhu

Sponsors

Parts

Basic Parts

BBa_K2346000 Profile

Nitric oxide reductase (NOR)

Coding sequence for enzyme that will catalyst the reduction of NO(nitrogen monoxide) to N2O (nitrous oxide).

BBa_K2346001 Profile

Nitrite reductase (NiR)

Coding sequence for enzyme that will catalyst the reduction of Nitrite ion (NO2-) to NO(nitrogen monoxide).

BBa_K2346002 Profile

Nitrous oxide reductase (nosZ)

Coding sequence for enzyme that will catalyst the reduction of N2O(nitrous oxide) to Nitrogen and Oxygen.

BBa_K2346009 Profile

GGGGS x2 Linker

Coding sequence for a flexible linker, it has advantages of improving protein solubility, providing enzyme flexibility for catalysis, domain separation.

BBa_K2346010 Profile

EAAAK x2 linker

Coding sequence for an α-helix forming linker, it has advantages of improving protein soluility, providing enzyme flexibility for catalysis Domain separation.

BBa_K2346011 Profile

Nitrite Degradation Fusion Enzyme(NzFE)

Coding sequence for a fusion enzyme made from nosZ, NiR, NOR. The start codons of NiR and NOR are eliminated during PCR stage of inFusion Assembly. The end codons of NiR and nosZ are eliminated during PCR stage of inFusion Assembly.

Composite Parts

BBa_K2346003 Profile

Nitrate/nitrite responsive promoter with amplifier

This is a composite part and a functional improved device for Part:BBa_K216005. The Pyear promoter will be switched on under the presence of nitrate or nitrite, translating P2 ogr. The P2 ogr will cause inducible PF promoter to produce even stronger downstream PoPS. We attached a RBS for convince of registry users.

BBa_K2346004 Profile

Nitrate/nitrite responsive promoter with autoinducer synthetase for AHL(3OC6HSL)

This is a composite part. The Pyear promoter will be switched on under the presence of nitrate or nitrite, translating LuxL protein. LuxL is an enzyme for catalyzing AHL(acyl-homoserine lactones) from normal cell metabolites.

BBa_K2346005 Profile

Positive Feedback Loop for NzFE

This is a device to reduce nitrite ion to its elements: oxygen and nitrogen. The device is regulated by signaling molecule AHL(3OC6HSL). The right p(tetR) promoter will constitutively express Plux's repressor: LuxR protein. Two LuxR protein molecules can bind with two AHL molecules to form a complex which in turns switches on the PluxR promoter. The down stream LuxL is an enzyme for catalyzing AHL(acyl-homoserine lactones) from normal cell metabolites. The LuxL translated will increase the AHL content in the chassis. This forms a positive feedback loop with increasing AHL resulting in increasing PoPS from PluxR. We utilize this increasing PoPS, and place CDS for NzFE(Nitrite Degradation Fusion Enzyme) in the down stream of the circuit.

BBa_K2346006 Profile

Enhanced Sensor for Nitrite/Nitrate

This is a functional improved device for BBa_K381001(http://parts.igem.org/Part:BBa_K381001). We replaced the original reporter gene GFP with brighter reporter gene sfGFP, in the hope to improve the sensitivity for the device to detect nitrite and nitrate.

BBa_K2346007 Profile

NzFE T7 expression vector

This is a composite part for expression of NzFE in DE-type bacteria. The pT7 promoter can only be used in chassis with T7 RNA synthase. Though pT7 commonly thought to be constitutive, we encountered the problem of unable to express the protein without IPTG (lactose replacement). We suspect the company providing BL21 DE strain competent cell changed the genome of their bacteria, made T7 RNA synthase regulated by IPTG.

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