LOCUS       Exported                7403 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPolyREP, complete sequence.
ACCESSION   KF680544
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  AUTHORS   Kolkenbrock S, Moerschbacher BM.
  TITLE     pPolyREPII, a Synthetic, Modular, Broad-Host-Range Expression Vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7403)
  AUTHORS   Kolkenbrock S, Moerschbacher BM.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-SEP-2013) Institute of Plant Biology and 
            Biotechnology, WWU Muenster, Schlossplatz 8, Muenster, NRW 48143, 
            Germany
REFERENCE   3  (bases 1 to 7403)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7403
                     /organism="Cloning vector pPolyREP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1435202"
     regulatory      123..193
                     /regulatory_class="promoter"
                     /note="Ptac promoter"
     promoter        125..153
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    161..177
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      269..296
                     /regulatory_class="terminator"
                     /note="trpA terminator"
     regulatory      305..332
                     /regulatory_class="terminator"
                     /note="trpA terminator"
     regulatory      333..362
                     /regulatory_class="terminator"
                     /note="t500 terminator"
     rep_origin      373..2550
                     /label=pWV01 replication origin
                     /note="pWV01 replication origin"
     CDS             complement(1083..1094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2559..2593
                     /regulatory_class="terminator"
                     /note="tR2 terminator"
     regulatory      2603..2636
                     /regulatory_class="terminator"
                     /note="tR2 terminator"
     CDS             complement(2654..3304)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol acetyltransferase
                     /protein_id="AHC08504.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     rep_origin      3496..5829
                     /label=pBBR1 replication origin
                     /note="pBBR1 replication origin"
     rep_origin      3790..4559
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             4560..5222
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     regulatory      5836..5879
                     /regulatory_class="terminator"
                     /note="rrnB terminator"
     terminator      5836..5879
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      6011..6038
                     /regulatory_class="terminator"
                     /note="rrnB terminator"
     terminator      6011..6038
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(6044..7126)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(6044..7003)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AHC08505.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     gene            complement(6044..7003)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(7127..7204)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      7292..7322
                     /regulatory_class="terminator"
                     /note="hisT terminator"
     regulatory      7349..7389
                     /regulatory_class="terminator"
                     /note="hisT terminator"