pMCSG20 载体 (V010705#)

载体基本信息
载体名称 pMCSG20 载体抗性 Ampicillin
载体长度 5991 bp 载体类型 Structural Genomics Vectors
载体来源 Eschenfeldt WH, Lucy S, Millard CS, Joachimiak A, Mark ID. 拷贝数 High copy number

pMCSG20 载体载体图谱

pMCSG205991 bp60012001800240030003600420048005400T7 terminator6xHisTEV siteGSTS-TagATGRBSlac operatorT7 promoterlacI promoterlacICAP binding siteropbomoriAmpRAmpR promoter

pMCSG20 载体载体序列

LOCUS       pMCSG20.        5991 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector with an S-Tag-GST-TEV leader for high-throughput 
            purification of recombinant proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pMCSG20.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5991)
  AUTHORS   Eschenfeldt WH, Lucy S, Millard CS, Joachimiak A, Mark ID.
  TITLE     A family of LIC vectors for high-throughput cloning and purification
            of proteins.
  JOURNAL   Methods Mol. Biol. 2009;498:105-15.
  PUBMED    18988021
REFERENCE   2  (bases 1 to 5991)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5991)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            Mol. Biol."; date: "2009"; volume: "498"; pages: "105-15"
COMMENT     SGRef: number: 2; 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..5991
                     /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(223..243)
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             complement(271..924)
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     CDS             complement(946..990)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(991..993)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(1001..1023)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1038..1062)
                     /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(1063..1081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1394..1471
                     /label=lacI promoter
     CDS             1472..2551
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2567..2588
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3363..3551
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3656..3798
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3984..4572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4746..5603)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5604..5707)
                     /label=AmpR promoter

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