pMCSG31 vector (V010683)

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LOCUS       pMCSG31.        5822 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial expression vector with an MBP-TVMV-6xHis-TEV leader and a 
            Tet-inducible TVMV protease cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pMCSG31
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5822)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5822)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For ligation-independent cloning (LIC), linearize with SspI and 
            treat with T4 DNA polymerase plus dGTP.
FEATURES             Location/Qualifiers
     source          1..5822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..91
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     RBS             130..152
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             184..891
                     /label=TVMV protease
                     /note="tobacco vein mottling virus NIa protease
                     (Nallamsetty et al., 2004)"
     terminator      927..1013
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        1244..1262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1263..1287
                     /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)."
     RBS             1318..1323
                     /note="ribosome binding site"
     CDS             1332..2432
                     /label=MBP
                     /note="maltose binding protein from E. coli"
     protein_bind    2442..2466
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2469..2489
                     /label=TVMV site
                     /note="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site"
     CDS             2490..2507
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2532..2552
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             2618..2635
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2702..2749
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             2818..2841
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     rep_origin      2932..3390
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3416..3519
                     /label=AmpR promoter
     CDS             3520..4377
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             4387..5010
                     /label=TetR
                     /note="tetracycline repressor TetR"
     rep_origin      5166..5754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"