pET-32 Xa/LIC vector (V010988)

Download GeneBank File(.gb)

LOCUS       pET-32_Xa_LIC.        5926 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for ligation-independent cloning (LIC) to express 
            thioredoxin-tagged proteins with a Factor Xa site.
ACCESSION   .
VERSION     .
KEYWORDS    pET-32 Xa_LIC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5926)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5926)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For LIC, linearize with BseRI and treat with T4 DNA polymerase plus 
            dCTP.
FEATURES             Location/Qualifiers
     source          1..5926
                     /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"
     misc_feature    158..216
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(233..244)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     CDS             complement(275..319)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(326..343)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(353..370)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(392..718)
                     /label=TrxA
                     /note="E. coli thioredoxin"
     RBS             complement(726..748)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(763..787)
                     /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(788..806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1119..1196
                     /label=lacI promoter
     CDS             1197..2276
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2292..2313
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3088..3276
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3381..3523
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3709..4297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4471..5328)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5329..5433)
                     /label=AmpR promoter
     rep_origin      complement(5460..5915)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"