pNH-TrxT vector (V010585)

LOCUS       40924_33187        7602 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNH-TrxT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  AUTHORS   Cooper CDO., Gileadi O.
  TITLE     
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7602)
  AUTHORS   Cooper CDO., Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2009) Structural Genomics Consortium, University 
            of Oxford, ORCRB, Old Road Campus, off Roosevelt Drive, Oxford, 
            Oxfordshire OX3 7DQ, United Kingdom
REFERENCE   3  (bases 1 to 7602)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7602)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-DEC-2009) Structural Genomics Consortium, University of Oxford, 
            ORCRB, Old Road Campus, off Roosevelt Drive, Oxford, Oxfordshire OX3
            7DQ, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             12..34
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             45..62
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             72..398
                     /label=TrxA
                     /note="E. coli thioredoxin"
     CDS             405..425
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     promoter        441..886
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             887..2305
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    2354..2368
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     CDS             2416..2433
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2500..2547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2584..3039
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(3135..3947)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4069..4657
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4843..4985)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5090..5278)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(6053..6074)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6090..7169)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7170..7247)
                     /label=lacI promoter
     promoter        7556..7574
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7575..7599
                     /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)."