pRH52 vector (V003739)

Basic Vector Information

Vector Name:
pRH52
Antibiotic Resistance:
Streptomycin
Length:
6389 bp
Type:
Cloning vector
Replication origin:
pBBR1 oriV
Source/Author:
Ghosh IN, Landick R.

pRH52 vector Map

pRH526389 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300lacIq promoterlacIB. subtilis glnA terminatorCAP binding siteTrc promoter (Ptrc); Trc promoter (Ptrc)lac operatorT3 promoterRBSsuperfolder GFPPhage 22 terminatorpBBR1 ReppBBR1 oriVAmpR promoterSmR

pRH52 vector Sequence

LOCUS       40924_36998        6389 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRH52, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6389)
  AUTHORS   Ghosh IN, Landick R.
  TITLE     OptSSeq: High-throughput sequencing readout of growth enrichment 
            defines optimal gene expression elements for homoethanologenesis
  JOURNAL   ACS Synth Biol (2016) In press
  PUBMED    27404024
REFERENCE   2  (bases 1 to 6389)
  AUTHORS   Ghosh IN, Landick RC.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2016) DOE Great Lakes Bioenergy Research Center, 
            University of Wisconsin-Madison, 1552 University Avenue, Madison, WI
            53726, United States
REFERENCE   3  (bases 1 to 6389)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6389)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JUL-2016) DOE Great Lakes Bioenergy Research Center, University 
            of Wisconsin-Madison, 1552 University Avenue, Madison, WI 53726, 
            United States"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     Cloning vector recieved with thanks from Haft, R.J.
FEATURES             Location/Qualifiers
     source          1..6389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        163..240
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             241..1320
                     /label=lacI
                     /note="lac repressor"
     regulatory      1326..1373
                     /label=B. subtilis glnA terminator
                     /note="B. subtilis glnA terminator"
                     /regulatory_class="terminator"
     protein_bind    complement(1385..1404)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    1404..1425
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     regulatory      1440..1468
                     /note="Trc promoter (Ptrc); Trc promoter (Ptrc)"
                     /regulatory_class="promoter"
     protein_bind    1477..1493
                     /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        1543..1561
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     RBS             1597..1619
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1627..2340
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     regulatory      2400..2451
                     /label=Phage 22 terminator
                     /note="Phage 22 terminator"
                     /regulatory_class="terminator"
     CDS             complement(2657..3316)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(3317..4086)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     promoter        5464..5555
                     /label=AmpR promoter
     CDS             5556..6344
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"

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