Team:Peking/Parts
Parts List
| Part Name | Part Type | Description |
|---|---|---|
| BBa_K3081055 | coding | dCas9-ssrA |
| BBa_K3081119 | coding | dCas9-linker-sfGFP |
| BBa_K3081122 | coding | dCas9-linker-sfGFP ssrA |
| BBa_K3081105 | RNA | control sgRNA |
| BBa_K3081106 | RNA | polyA |
| BBa_K3081107 | RNA | M+ |
| BBa_K3081108 | RNA | R1+ |
| BBa_K3081109 | RNA | R1+ (18bp) |
| BBa_K3081110 | RNA | R1+ (19bp) |
| BBa_K3081111 | RNA | R1- |
| BBa_K3081112 | RNA | R4- |
| BBa_K3081113 | RNA | MR13 |
| BBa_K3081114 | RNA | IHF |
| BBa_K3081115 | RNA | R3+ |
| BBa_K3081116 | RNA | M-R2+ |
| BBa_K3081120 | RNA | pL01 |
| BBa_K3081121 | RNA | pL05 |
| BBa_K3081130 | RNA | NT1 |
| BBa_K3081104 | RNA | gRNA scaffold |
| BBa_K3081056 | composite | J23119 sgRNA genarator |
| BBa_K3081002 | composite | pBAD-dCas9 linker sfGFP |
| BBa_K3081003 | composite | pBAD-dCas9 linker sfGFP ssrA |
| BBa_K3081004 | composite | pTAC-dCas9 linker GFP |
| BBa_K3081060 | composite | pBAD-dCas9-T7-M+ |
| BBa_K3081058 | composite | pBAD-dCas9-J23119-sgRNA |
| BBa_K3081006 | composite | pBAD-dCas9-J23119-polyA |
| BBa_K3081007 | composite | pBAD-dCas9-J23119-M+ |
| BBa_K3081008 | composite | pBAD-dCas9-J23119-R1+ |
| BBa_K3081009 | composite | pBAD-dCas9-J23119-R1+(18bp) |
| BBa_K3081010 | composite | pBAD-dCas9-J23119-R1+(19bp) |
| BBa_K3081011 | composite | pBAD-dCas9-J23119-R1- |
| BBa_K3081012 | composite | pBAD-dCas9-J23119-R4- |
| BBa_K3081013 | composite | pBAD-dCas9-J23119-MR13 |
| BBa_K3081014 | composite | pBAD-dCas9-J23119-IHF |
| BBa_K3081015 | composite | pBAD-dCas9-J23119-R3+ |
| BBa_K3081016 | composite | pBAD-dCas9-J23119-M-R2+ |
| BBa_K3081022 | composite | pBAD-dCas9-J23119-NT1 |
| BBa_K3081062 | composite | pBAD-dCas9-J23119-pL01 |
| BBa_K3081063 | composite | pBAD-dCas9-J23119-pL05 |
| BBa_K3081024 | composite | pBAD-dCas9-J23119-NT1-J23119-mRFP |
| BBa_K3081027 | composite | pBAD-dCas9-J23119-M+-J23119-R1+ |
| BBa_K3081031 | composite | pBAD-dCas9-ssrA-J23119-R1+ |
| BBa_K3081032 | composite | pBAD-dCas9-ssrA-J23119-R1+(18bp) |
| BBa_K3081033 | composite | pBAD-dCas9-ssrA-J23119-R1+(19bp) |
| BBa_K3081057 | composite | pBAD-dCas9-ssrA-J23119-p15A-initiation(+) |
| BBa_K3081041 | composite | pBAD-dCas9-ssrA-J23119-p15A-initiation(-) |
| BBa_K3081042 | composite | pBAD-dCas9-ssrA-J23119-p15A-RNA II NT |
| BBa_K3081043 | composite | pBAD-dCas9-ssrA-J23119-p15A-RNA I NT |
| BBa_K3081044 | composite | pBAD-dCas9-ssrA-J23119-NT1-J23119-mRFP |
| BBa_K3081046 | composite | pBAD-dCas9-T7-sgRNA |
| BBa_K3081053 | composite | ptet-luxR-plux-dCas9-T7-sgRNA |
| BBa_K3081054 | composite | ptet-luxR-plux-dCas9-T7-M+ |
| BBa_K3081051 | composite | lacI-plac-luxI-luxR-plux-GFP |
| BBa_K3081052 | composite | ptet-luxR-plux-GFP-B1002 |
| BBa_K3081053 | composite | ptet-luxR-plux-dCas9-T7-M+ |
| BBa_K3081054 | composite | ptet-luxR-plux-dCas9-T7-sgRNA |
| BBa_K3081059 | composite | TcI42-dCas9-J23119-M+ |
| BBa_K3081061 | composite | TcI42-dCas9-J23119-polyA |
Part Collection
2019 Peking iGEM constructed a large part collection focusing on dCas9-based DNA replication control. The dCas9 expression module has been optimized and expanded by using different regulatory elements, adding degradation tag, fusing a fluorescent protein reporter, etc. The updated modules enable our system to function in a low-leakage, quantifiable and reversible manner, and a series of application scenarios. We also designed a new expression module for sgRNA. The new expression system of two inputs not only benefit our project but also can be a new paradigm of CRISPRi for subsequent researchers. We have also constructed a library of sgRNA to target the replication origin of E. coli genome or plasmids (p15A and pSC101). sgRNAs are different in target sites, length and number, thus can perform a wide range of control effects. All the parts are thoroughly characterized by different methods: microscopic imaging, spectrophotometer, cytometer, microfluidic devices, qPCR, etc.