LOCUS       Exported                4041 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  UNVERIFIED: Cloning vector pSEVA-Plac-E2-Orange, complete sequence.
ACCESSION   MH013365
VERSION     .
KEYWORDS    UNVERIFIED.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4041)
  AUTHORS   Zhang C, Parrello D, Brown PJ, Wall JD, Hu Z.
  TITLE     A novel whole-cell biosensor of Pseudomonas aeruginosa to monitor 
            the expression of quorum sensing genes
  JOURNAL   Appl. Microbiol. Biotechnol. (2018) In press
  PUBMED    29730766
REFERENCE   2  (bases 1 to 4041)
  AUTHORS   Zhang C.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-FEB-2018) Department of Civil & Environmental 
            Engineering, University of Missouri-Columbia, E2509 Lafferre Hall, 
            Columbia, MO 65211, USA
REFERENCE   3  (bases 1 to 4041)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     GenBank staff is unable to verify sequence and/or annotation 
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..4041
                     /organism="Cloning vector pSEVA-Plac-E2-Orange"
                     /mol_type="other DNA"
                     /note="plasmid constructed by Patrick Billard (Universite 
                     de Lorraine, CNRS, Laboratoire Interdisciplinaire des 
                     Environnements Continentaux (LIEC), UMR 7360, 
                     Vandoeuvre-les-Nancy, France)"
                     /db_xref="taxon:2201086"
     protein_bind    282..298
                     /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)."
     protein_bind    315..331
                     /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)."
     CDS             385..1062
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKTIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     terminator      1073..1167
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1299..2114
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2281..2389
                     /note="incP origin of transfer"
     CDS             complement(2400..3062)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3063..3834
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      3949..4035
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"