Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Erik Lundin
Date created: 2012-09-22 11:00:00
Date modified: 2015-05-08 01:13:37
Low copy BioBrick standard vector
| Types | DnaRegion |
| Roles | Plasmid_Backbone plasmid_vector |
| Sequences | BBa_K864005_sequence (Version 1) |
Description
pSB4A15 is a BioBrick standard vector with low copy pSC101 replication origin (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.The resistance cassette is flanked by FRT sites (BBa_J61020) which allows for subsequent removal
of the resistance cassette from the chromosome by the FLP recombinase. The pSB8x15 series are especially suitable for carrying the FLP recombinase, since they can be removed by incubation at 42??C.
Notes
The resistance cassette is terminated by a Lux bidirectional terminator. Adjacent to this terminator there is a designed primer binding site to where the forward primer for lambda red can be designed to bind to no matter what the insert in the cloning site is or what the resistance is.The resistance cassette is flanked by SalI and SacI restriction sites for easy switching of resistance in the backbone.
The pSC101 origin of replication is flaked by NheI and MluI restriction sites for easy switching of origin of replication.
Source
Based on pSB3T5| Sequence Annotation | Location | Component / Role(s) |
| Lambda Red reverse primer TonB terminator VR rrnC terminator pSC101 ori BBa_K864050 Lambda Red forward primer Lux terminator | 18,38 47,115 116,135 136,175 190,2405 2433,2450 2454,2499 | primer_binding_site feature/primer_binding feature/stem_loop stem_loop feature/primer_binding primer_binding_site stem_loop feature/stem_loop sequence_feature feature/misc feature/primer_binding primer_binding_site feature/stem_loop stem_loop |