LOCUS       40924_39183        4041 bp DNA     circular SYN 18-DEC-2018
DEFINITION  UNVERIFIED: Cloning vector pSEVA-Plac-E2-Orange, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
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   
REFERENCE   3  (bases 1 to 4041)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4041)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-FEB-2018) Department of Civil "
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     GenBank staff is unable to verify sequence and/or annotation 
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..4041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    282..298
                     /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)."
     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..1059
                     /codon_start=1
                     /label=DsRed-Express2
                     /note="noncytotoxic tetrameric variant of DsRed fluorescent
                     protein (Strack et al., 2008)"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKTIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     terminator      1073..1167
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1299..2111
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2281..2389
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2403..3062)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(3063..3834)
                     /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"
     terminator      complement(3949..4035)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt accatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      121 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
      181 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
      241 tcgtcttcac ctcgagaaaa tttatcaaaa agagtgttga cttgtgagcg gataacaatg
      301 atacttagat tcaattgtga gcggataaca atttcacaca tctagagtcg acctgcaggc
      361 atgcaagctt aggaggaaaa acatatggat agcactgaga acgtcatcaa gcccttcatg
      421 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
      481 ggcgagggca agccctacga gggcacccag accgccaagc tgcaggtgac caagggcggc
      541 cccctgccct tcgcctggga catcctgtcc ccccagttca cctacggctc caaggcctac
      601 gtgaagcacc ccgccgacat ccccgactac aagaagctgt ccttccccga gggcttcaag
      661 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
      721 ctgcaggacg gcaccttcat ctaccacgtg aagttcatcg gcgtgaactt cccctccgac
      781 ggccccgtaa tgcagaagaa gactctgggc tgggagccct ccaccgagcg cctgtacccc
      841 cgcgacggcg tgctgaaggg cgagatccac aaggcgctga agctgaaggg cggcggccac
      901 tacctggtgg agttcaagac aatctacatg gccaagaagc ccgtgaagct gcccggctac
      961 tactacgtgg actccaagct ggacatcacc tcccacaacg aggactacac cgtggtggag
     1021 cagtacgagc gcgccgaggc ccgccaccac ctgttccagt agactagtct tggactcctg
     1081 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1141 cgccgggcgt tttttattgg tgagaatcca ggggtcccca ataattacga tttaaatttg
     1201 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1261 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa
     1321 cgagctgtag ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat
     1381 gggcgcgtga taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg
     1441 atgcgccgga actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg
     1501 aaatggtgcg tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta
     1561 ttcgcacccc ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc
     1621 aggtgctgga agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc
     1681 tgcgtcgtct gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc
     1741 gtctggccca ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg
     1801 atgaacgtaa cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt
     1861 ttagcccgga tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg
     1921 aaggcaaact gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg
     1981 atctggccat tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt
     2041 ttcagaaata tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg
     2101 atgaattttt ctaataatta attggaccgc ggtccgcgcg ttgtcctttt ccgctgcata
     2161 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
     2221 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
     2281 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
     2341 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgt aggccggccc
     2401 taccggcgcg gcagcgttac ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa
     2461 cacggctcat cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg
     2521 gtcaaggctg gcaggtctgg ttccatgccc ggaatgccgg gctggctggg cggctcctcg
     2581 ccggggccgg tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg
     2641 ccatgcccca acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac
     2701 accgacaggc gcaactggtc gcggggctgg ccccacgcca cgcggtcatt gaccacgtag
     2761 gccgacacgg tgccggggcc gttgagcttc acgacggaga tccagcgctc ggccaccaag
     2821 tccttgactg cgtattggac cgtccgcaaa gaacgtccga tgagcttgga aagtgtcttc
     2881 tggctgacca ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg cagcagcatt
     2941 gccgccgtgg gtttcctcgc aataagcccg gcccacgcct catgcgcttt gcgttccgtt
     3001 tgcacccagt gaccgggctt gttcttggct tgaatgccga tttctctgga ctgcgtggcc
     3061 atgcttatct ccatgcggta ggggtgccgc acggttgcgg caccatgcgc aatcagctgc
     3121 aacttttcgg cagcgcgaca acaattatgc gttgcgtaaa agtggcagtc aattacagat
     3181 tttctttaac ctacgcaatg agctattgcg gggggtgccg caatgagctg ttgcgtaccc
     3241 ccctttttta agttgttgat ttttaagtct ttcgcatttc gccctatatc tagttctttg
     3301 gtgcccaaag aagggcaccc ctgcggggtt cccccacgcc ttcggcgcgg ctccccctcc
     3361 ggcaaaaagt ggcccctccg gggcttgttg atcgactgcg cggccttcgg ccttgcccaa
     3421 ggtggcgctg cccccttgga acccccgcac tcgccgccgt gaggctcggg gggcaggcgg
     3481 gcgggcttcg cccttcgact gcccccactc gcataggctt gggtcgttcc aggcgcgtca
     3541 aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc gccttccact tggtgtccaa
     3601 ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt ccccagagaa aattaaaaaa
     3661 attgatgggg caaggccgca ggccgcgcag ttggagccgg tgggtatgtg gtcgaaggct
     3721 gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc aggcgcagcc tgtccatcag
     3781 cttgtccagc agggttgtcc acgggccgag cgaagcgagc cagccggtgg ccgctcgcgg
     3841 ccatcgtcca catatccacg ggctggcaag ggagcgcagc gaccgcgcag ggcgaagccc
     3901 ggagagcaag cccgtagggg gggcgcgccc agctgtctag ggcggcggat ttgtcctact
     3961 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
     4021 ctttcgtttt atttgatgcc t
//
