pVP13 vector (V010573)

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LOCUS       pVP13.        7675 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial Gateway(R) destination vector for appending an N-terminal 
            S-Tag-6xHis-MBP cassette to an expressed protein. Also known as 
            pVP13-GW.
ACCESSION   .
VERSION     .
KEYWORDS    pVP13
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7675)
  AUTHORS   Thao S, Zhao Q, Kimball T, Steffen E, Blommel PG, Riters M, Newman 
            CS, Fox BG, Wrobel RL.
  TITLE     Results from high-throughput DNA cloning of Arabidopsis thaliana 
            target genes using site-specific recombination.
  JOURNAL   J. Struct. Funct. Genomics 2004;5:267-76.
  PUBMED    15750721
REFERENCE   2  (bases 1 to 7675)
  AUTHORS   Center for Eukaryotic Structural Genomics
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7675)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Struct. 
            Funct. Genomics"; date: "2004"; volume: "5"; pages: "267-76"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     A TEV cleavage site can be encoded during PCR amplification.
FEATURES             Location/Qualifiers
     source          1..7675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..55
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    63..79
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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             102..107
                     /note="ribosome binding site"
     CDS             116..118
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             122..166
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             173..190
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             194..1291
                     /label=MBP
                     /note="maltose binding protein from E. coli"
     protein_bind    1361..1485
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1510..1540
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1594..2250
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             2595..2897
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2941..3065)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3133..3227
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3271..3927
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      3995..4081
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(4140..4161)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4177..5256)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5257..5334)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    5526..5666
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5852..6440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6614..7471)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7472..7576)
                     /label=AmpR promoter