pMCSG61 vector (V010625)

MSDS

Basic Vector Information

Vector Name:: pMCSG61

Antibiotic Resistance:: Ampicillin

Length:: 6422 bp

Type:: Structural Genomics Vectors

Replication origin:: ori

Source/Author:: Midwest Center for Structural Genomics

Copy Number:: High copy number

pMCSG61 vector Vector Map

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Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pMCSG61 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pMCSG61.        6422 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial expression vector with a 6xHis-TEV-MBP leader, for 
            high-throughput purification of recombinant proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pMCSG61.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6422)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For ligation-independent cloning (LIC), amplify with LIC-F and LIC-R
            and treat with T4 DNA polymerase plus dTTP.
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(256..1353)
                     /label=MBP
                     /note="maltose binding protein from E. coli"
     CDS             complement(1360..1380)
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             complement(1405..1422)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(1423..1425)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(1433..1455)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1469..1493)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1494..1512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1825..1902
                     /label=lacI promoter
     CDS             1903..2982
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2998..3019
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3794..3982
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4087..4229
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4415..5003)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5177..6034)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6035..6138)
                     /label=AmpR promoter

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