Basic Vector Information
- Vector Name:
- pMCSG35
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7484 bp
- Type:
- Structural Genomics Vectors
- Replication origin:
- ori
- Source/Author:
- Eschenfeldt WH, Maltseva N, Stols L, Donnelly MI, Gu M, Nocek B, Tan
- Copy Number:
- High copy number
pMCSG35 vector Map
pMCSG35 vector Sequence
LOCUS pMCSG35. 7484 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial vector encoding N-terminal 10xHis-MBP-TEV and C-terminal TEV-6xHis plus TVMV protease. See also pMCSG35B. ACCESSION . VERSION . KEYWORDS pMCSG35 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7484) AUTHORS Eschenfeldt WH, Maltseva N, Stols L, Donnelly MI, Gu M, Nocek B, Tan K, Kim Y, Joachimiak A. TITLE Cleavable C-terminal His-tag vectors for structure determination. JOURNAL J. Struct. Funct. Genomics 2010;11:31-9. PUBMED 20213425 REFERENCE 2 (bases 1 to 7484) AUTHORS Midwest Center for Structural Genomics TITLE Direct Submission REFERENCE 3 (bases 1 to 7484) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct. Funct. Genomics"; date: "2010"; volume: "11"; pages: "31-9" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT For ligation-independent cloning (LIC), linearize with SmaI and treat with T4 DNA polymerase plus dATP. FEATURES Location/Qualifiers source 1..7484 /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(208..225) /label=6xHis /note="6xHis affinity tag" CDS complement(232..252) /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" CDS complement(263..283) /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" CDS complement(305..1402) /label=MBP /note="maltose binding protein from E. coli" CDS complement(1430..1456) /label=9xHis /note="9xHis affinity tag" CDS complement(1457..1459) /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" RBS complement(1467..1489) /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" protein_bind complement(1504..1528) /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(1529..1547) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 1846..1919 /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" CDS 1943..2650 /label=TVMV protease /note="tobacco vein mottling virus NIa protease (Nallamsetty et al., 2004)" terminator 2686..2772 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" promoter 2887..2964 /label=lacI promoter CDS 2965..4044 /label=lacI /note="lac repressor" protein_bind 4060..4081 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS 4856..5044 /label=rop /note="Rop protein, which maintains plasmids at low copy number" misc_feature 5149..5291 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(5477..6065) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6239..7096) /label=AmpR /note="beta-lactamase" promoter complement(7097..7200) /label=AmpR promoter
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