LOCUS       40924_39193        4062 bp DNA     circular SYN 18-DEC-2018
DEFINITION  UNVERIFIED: Cloning vector pSEVA-Plac-mTag BFP2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4062)
  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 4062)
  AUTHORS   Zhang C.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 4062)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4062)
  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..4062
                     /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..1083
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     terminator      1094..1188
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1320..2132
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2302..2410
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2424..3083)
                     /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(3084..3855)
                     /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(3970..4056)
                     /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 acatatgagc gagctgatta aggagaacat gcacatgaag
      421 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
      481 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
      541 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
      601 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
      661 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      721 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
      781 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
      841 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
      901 aacgccaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
      961 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag
     1021 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt
     1081 aatactagtc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     1141 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggggtcccc
     1201 aataattacg atttaaattt gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1261 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1321 tgagccatat tcagcgtgaa acgagctgta gccgtccgcg tctgaacagc aacatggatg
     1381 cggatctgta tggctataaa tgggcgcgtg ataacgtggg tcagagcggc gcgaccattt
     1441 atcgtctgta tggcaaaccg gatgcgccgg aactgtttct gaaacatggc aaaggcagcg
     1501 tggcgaacga tgtgaccgat gaaatggtgc gtctgaactg gctgaccgaa tttatgccgc
     1561 tgccgaccat taaacatttt attcgcaccc cggatgatgc gtggctgctg accaccgcga
     1621 ttccgggcaa aaccgcgttt caggtgctgg aagaatatcc ggatagcggc gaaaacattg
     1681 tggatgcgct ggccgtgttt ctgcgtcgtc tgcatagcat tccggtgtgc aactgcccgt
     1741 ttaacagcga tcgtgtgttt cgtctggccc aggcgcagag ccgtatgaac aacggcctgg
     1801 tggatgcgag cgattttgat gatgaacgta acggctggcc ggtggaacag gtgtggaaag
     1861 aaatgcataa actgctgccg tttagcccgg atagcgtggt gacccacggc gattttagcc
     1921 tggataacct gattttcgat gaaggcaaac tgattggctg cattgatgtg ggccgtgtgg
     1981 gcattgcgga tcgttatcag gatctggcca ttctgtggaa ctgcctgggc gaatttagcc
     2041 cgagcctgca aaaacgtctg tttcagaaat atggcattga taatccggat atgaacaaac
     2101 tgcaatttca tctgatgctg gatgaatttt tctaataatt aattggaccg cggtccgcgc
     2161 gttgtccttt tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata
     2221 tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg
     2281 gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg
     2341 ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg
     2401 aggctggccg taggccggcc ctaccggcgc ggcagcgtta cccgtgtcgg cggctccaac
     2461 ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc
     2521 cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg
     2581 ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc cggatacagg
     2641 gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca
     2701 accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg gccccacgcc
     2761 acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt cacgacggag
     2821 atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa agaacgtccg
     2881 atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc catctgcgcc
     2941 acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc ggcccacgcc
     3001 tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc ttgaatgccg
     3061 atttctctgg actgcgtggc catgcttatc tccatgcggt aggggtgccg cacggttgcg
     3121 gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg cgttgcgtaa
     3181 aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc ggggggtgcc
     3241 gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc tttcgcattt
     3301 cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc
     3361 cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc
     3421 gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca ctcgccgccg
     3481 tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac tgcccccact cgcataggct
     3541 tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc
     3601 cgccttccac ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc cggaggcttt
     3661 tccccagaga aaattaaaaa aattgatggg gcaaggccgc aggccgcgca gttggagccg
     3721 gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca agctcgtggg
     3781 caggcgcagc ctgtccatca gcttgtccag cagggttgtc cacgggccga gcgaagcgag
     3841 ccagccggtg gccgctcgcg gccatcgtcc acatatccac gggctggcaa gggagcgcag
     3901 cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg ggggcgcgcc cagctgtcta
     3961 gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     4021 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ct
//
