pNIC-Bio2 vector (V004323)

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LOCUS       40924_33207        7335 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNIC-Bio2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7335)
  AUTHORS   Keates T, Cooper CD, Savitsky P, Allerston CK, Phillips C, 
            Hammarstrom M, Daga N, Berridge G, Mahajan P, Burgess-Brown NA, 
            Muller S, Graslund S, Gileadi O.
  TITLE     Expressing the human proteome for affinity proteomics: optimising 
            expression of soluble protein domains and in vivo biotinylation
  JOURNAL   N Biotechnol 29 (5), 515-525 (2012)
  PUBMED    22027370
REFERENCE   2  (bases 1 to 7335)
  AUTHORS   Graslund S, Hammarstrom M, Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAY-2011) Structural Genomics Consortium, SGC, 
            University of Oxford, ORCRB, Roosevelt Drive, Oxford OX3 7DQ, UK
REFERENCE   3  (bases 1 to 7335)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7335)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "N 
            Biotechnol"; date: "2012"; volume: "29"; issue: "5"; pages: 
            "515-525"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-MAY-2011) Structural Genomics Consortium, SGC, University of 
            Oxford, ORCRB, Roosevelt Drive, Oxford OX3 7DQ, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..118
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             134..154
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     promoter        170..615
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             616..2034
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    complement(2084..2097)
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     CDS             2099..2143
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     CDS             2196..2213
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2280..2327
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2364..2819
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2915..3727)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3849..4437
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4623..4765)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4870..5058)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5833..5854)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5870..6949)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6950..7027)
                     /label=lacI promoter