pEU-HSBC vector (V010745)

LOCUS       pEU-HSBC.        6336 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Flexi(R) Vector with the sacB gene, for in vitro transcription and 
            translation of an N-terminally 6xHis-tagged protein.
ACCESSION   .
VERSION     .
KEYWORDS    pEU-HSBC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  AUTHORS   Vinarov DA, Lytle BL, Peterson FC, Tyler EM, Volkman BF, Markley JL.
  TITLE     Cell-free protein production and labeling protocol for NMR-based 
            structural proteomics.
  JOURNAL   Nat. Methods 2004;1:149-53.
  PUBMED    15782178
REFERENCE   2  (bases 1 to 6336)
  AUTHORS   Center for Eukaryotic Structural Genomics
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6336)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2004"; volume: "1"; pages: "149-53"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     To remove the sacB gene, insert an ORF between the SgfI site and the
            PmeI site or a downstream site.
FEATURES             Location/Qualifiers
     source          1..6336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        373..391
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             521..523
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             527..544
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             790..2208
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        2297..2327
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2381..3037
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     misc_feature    3050..3105
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3795..3811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3819..3835)
                     /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(3843..3873)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3888..3909)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4197..4785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4959..5816)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5817..5921)
                     /label=AmpR promoter