Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: iGEM11_UNAM-Genomics_ Mexico
Date created: 2011-09-24 11:00:00
Date modified: 2015-05-08 01:12:36
pBBR1MCS-5 -> broad-host-range cloning vector
| Types | DnaRegion |
| Roles | plasmid_vector Plasmid_Backbone |
| Sequences | BBa_K535007_sequence (Version 1) |
Description
The plasmid we designed is a modification of the pBBR1MCS-5 , this broad-host-range (bhr) vector has been designed to assist the genetic analysis in prokaryotes, specifically in Gram- bacteria that are naturally CmR (chloramphenicol resistant).The pBBR1MCS-5 vector is relatively small (4768bp), it has an extended multiple cloning site (MCS) with 15 different restriction sites and it possesses a gentamicin resistance gene. After modification, only 3 of the 15 retrictions sites are present (see design notes).
The plasmid is mobilizable when the RK2 transfer functions is provided in trans and is compatible with IncP, IncQ and IncW group plasmids, as well as with ColE1- and P15a-based replicons.
We used this plasmid to transfer genes from E. coli to R. etli. First we transformed E. coli S17 with this vector containing the genes or sequences that we wanted to transfer, then we did a conjugation between our transformed E. coli cells and Rhizobium etli CFN42
Notes
It was amplified the BBa_J04450 including the prefix and the suffix from the psb1t3 plasmid and adding apaI and sacI restriction sites at the ends of the sequence. After, a ligation by sticky ends was with the pBBR1MCS-5 the apaI and sacI enzimes, trying to eliminate as mouch as possible of the MCS??? restriction sitesWe use the sequence as reported by Kovach et al. (1995).
Source
This is one of the antibiotic-resistant derivatives of the broad-host-range cloning vector pBBR1MCS.| Sequence Annotation | Location | Component / Role(s) |
| BioBrick suffix E coli his operon terminator stem loop sacI restriction site REP Gentamicine resistance marker apaI restriction site end of terminator stem loop BioBrick prefix | 1,21 22,93 30,65 232,237 997,1251 3712,3966 4904,4909 4963,4967 4977,4996 5006,5027 | sequence_feature feature/dna feature/stem_loop stem_loop stem_loop feature/stem_loop sequence_feature feature/misc feature/cds CDS CDS feature/cds feature/misc sequence_feature feature/stem_loop stem_loop stem_loop feature/stem_loop feature/dna sequence_feature |