pH3c-LIC vector (V010741)

LOCUS       pH3c-LIC.        5387 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expressing a protein with an N-terminal 8xHis 
            tag followed by an HRV 3C protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    pH3c-LIC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  AUTHORS   Cormier CY, Mohr SE, Zuo D, Hu Y, Rolfs A, Kramer J, Taycher E, 
            Kelley F, Fiacco M, Turnbull G, LaBaer J.
  TITLE     Protein Structure Initiative Material Repository: an open shared 
            public resource of structural genomics plasmids for the biological 
            community.
  JOURNAL   Nucleic Acids Res. 2010;38:D743-9.
  PUBMED    19906724
REFERENCE   2  (bases 1 to 5387)
  AUTHORS   Center for Structures of Membrane Proteins
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5387)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2010"; volume: "38"; pages: "D743-9"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     For ligation-independent cloning (LIC), linearize with SmaI and 
            treat with T4 DNA polymerase plus dTTP.
FEATURES             Location/Qualifiers
     source          1..5387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2271..2413)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label=lacI promoter
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /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             5042..5064
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5072..5074
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             5075..5098
                     /label=8xHis
                     /note="8xHis affinity tag"
     CDS             5132..5155
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             5168..5185
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             5231..5248
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      5315..5362
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"