Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Xinyu Liu
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-13 09:28:13
P(3HB)(BBa_K934001) + HSP promotor (BBa_K873002)
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1891014_sequence (Version 1) |
Description
Poly-3-hydroxybutyrate, P(3HB), a kind of PHAs is synthesized by three enzymes.The A gene encodes for the 393 amino acids protein, 3-ketothiolase (PhaA). The B1 gene encodes for the 246 amino acids protein, acetoacetyl-CoA reductase (PhaB). The C1 gene encodes for the 589 amino acids protein, PHA Synthase (PhaC)These three enzymes can use Co-A as a substrate to produce P(3HB). IGEM team Tokyo Tech2012 designed and synthesized phaC1-A-B1gene sequence BBa_K934001, and expressed via constitutive promoter.
In order to make the production process more controllable, we added arabinose inducible promoter（BBa_I0500）to phaC1-A-B1 gene sequence (BBa_K934001), constructing the arabinose inducible parts(BBa_K1891014）. Finally we linked the newly synthesized part to the standard vector pSB1C3. The new plasmids can be used directly for arabinose induced expression in E.coli.
Notes
We used Spe I and Pst I to do dual digestion in PSB1C3 which contained pBAD gene and recycled gene fragments in 3280bp, used Xbal I and Pst I to do dual digestion in PSB1C3 and recycled gene fragments in 4208bp.We use T4 ligase to link phaC1-A-B1 to the downstream of pBAD promoter, then transform the vectors to Trans 5α for plasmid extraction.Source
We get parts BBa_K934001 and BBa_I0500 from the kits sent by iGEM HQ.| Access | Instance | Definition |
| public public | BBa_I0500 BBa_K934001 | pBad/araC phaC1-A-B1 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_I0500 BBa_K934001 | 1,1210 1219,5426 | pBad/araC phaC1-A-B1 |