LOCUS       V001748                 9409 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001748
VERSION     V001748
KEYWORDS    pMo130-TelR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9409)
  AUTHORS   Amin IM, Richmond GE, Sen P, Koh TH, Piddock LJ, Chua KL
  TITLE     A method for generating marker-less gene deletions in
            multidrug-resistant Acinetobacter baumannii.
  JOURNAL   BMC Microbiol. 2013 Jul 13;13:158. doi: 10.1186/1471-2180-13-158.
   PUBMED   23848834
REFERENCE   2  (bases 1 to 9409)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9409)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC
            Microbiol."; date: "2013-07-13"; pages: "
            10.1186/1471-2180-13-158"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(972..1003)
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     protein_bind    complement(1067..1114)
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      complement(1293..1881)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            complement(2181..2290)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     protein_bind    complement(3501..3522)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     terminator      3540..3583
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3600..4952)
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
     protein_bind    complement(5078..5094)
                     /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)."
     CDS             complement(5156..5947)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(5948..6051)
                     /label="AmpR promoter"
     primer_bind     6119..6137
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(6175..6197)
                     /label="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6288..7208)
                     /gene="xylE"
                     /label="Metapyrocatechase"
                     /note="Metapyrocatechase from Pseudomonas putida.
                     Accession#: P06622"
     promoter        complement(7240..7268)
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    complement(7319..7366)
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     RBS             7458..7480
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atgaatgcca caaacaccga tgttttcgcc caggtaggcg gcctggaggc ccgaggcgcg
       61 aagatgaaga agcggggcac ccgcttcctc atcgcggcgc tggcagtcct tgccattgcc
      121 gggatcgggg cagtaacggg atgggcgatc agcccgagcg cgacgcccgg aagcattgac
      181 gtgccgcagg tgctggcatc gacattcagc gaccaggtgc cgggcagtga gggcggcggc
      241 ctgggtggcg gcctgccctt cacttcggcc gtcggggcat tcacggactt catggcgggg
      301 ccggcaattt ttaccttggg cattcttggc atagtggtcg cgggtgccgt gctcgtgttc
      361 gggggtgagt tctgcgggtt cgtgcgatcc gtctgcatga tggtgatagc cgtcagcatg
      421 attttcgtgt cgtcgaactt ggtgaagggc attctcggcg gcgatcacga cgccggccct
      481 gcggagcctt cgccgcgtgc gcgattcatg gcggccgtgg aggccaagga tttcgcgcga
      541 gtgcaagagc tgatcgaggc gcgtggagcc aagtcggcgg ctgattatgt ccttgcgcag
      601 ctcgccgtgg ccgaaggtct ggaccgcaag cctggtgcgc gcgtcgtggt cgggaaagcg
      661 gcgggcagca tggcaatgcc gcctgcggcg ctgggtttta cgccaagggg agaagcggca
      721 tacgccatcg agcggtcagc ctatggtgag ccgaggtcca gcattgcgaa gcagtaccag
      781 caggaatgga accggaaggc ggcgacctgg tgggcgatgg ccggtgtggc cggcatcatc
      841 ggcgcgatcc tggcggcggc ggcaaccggc tttgttgggc tggcagtgtc gatccgcaac
      901 cgagtgaagc gcgtgcgcga cctgttggtg atggagccgg gtgcagagcc ataagggatc
      961 gatccccctc aagtcaaaag cctccggtcg gaggcttttg actttctgct cgaggagcta
     1021 attccattga gtaagttttt aagcacatca gcttcaaaag cgctctgaag ttcctatact
     1081 ttctagagaa taggaacttc ggaataggta cttcaagatc cccaattcga gctcggtacc
     1141 cgggaagatc tggctagcgg ggccctgata tcagctcact caaaggcggt aatacggtta
     1201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2041 tggtctgaca gttattaatt aatcctcaat cgctcttcgt tcgtctggaa ggcagtacac
     2101 cttgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc
     2161 atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc
     2221 caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc
     2281 tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca
     2341 aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc
     2401 gaagcagggt tatgcagcgg aaaagcgctg cttccctgct gttttgtgga atatctaccg
     2461 actggaaaca ggcaaatgca ggaaattact gaactgaggg gacaggcgag agacgatgcc
     2521 aaagagctac accgacgagc tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg
     2581 gcgtgatgag gctgcggttg cgttcctccg cggcagatgt ctgcctgttc atccgcgtcc
     2641 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     2701 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     2761 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     2821 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     2881 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     2941 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     3001 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     3061 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     3121 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     3181 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     3241 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     3301 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     3361 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     3421 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3481 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgga attctcagat
     3541 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgaaag ctactattat
     3601 ttgttaactg ttaattgtcc ttgttcaagg atgctgtctt tgacaacaga tgttttcttg
     3661 cctttgatgt tcagcaggaa gctcggcgca aacgttgatt gtttgtctgc gtagaatcct
     3721 ctgtttgtca tatagcttgt aatcacgaca ttgtttcctt tcgcttgagg tacagcgaag
     3781 tgtgagtaag taaaggttac atcgttagga tcaagatcca tttttaacac aaggccagtt
     3841 ttgttcagcg gcttgtatgg gccagttaaa gaattagaaa cataaccaag catgtaaata
     3901 tcgttagacg taatgccgtc aatcgtcatt tttgatccgc gggagtcagt gaacaggtac
     3961 catttgccgt tcattttaaa gacgttcgcg cgttcaattt catctgttac tgtgttagat
     4021 gcaatcagcg gtttcatcac ttttttcagt gtgtaatcat cgtttagctc aatcataccg
     4081 agagcgccgt ttgctaactc agccgggcgt tttttatcgc tttgcagaag tttttgactt
     4141 tcttgacgga agaatgatgt gcttttgcca tagtatgctt tgttaaataa agattcttcg
     4201 ccttggtagc catcttcagt tccagtgttt gcttcaaata ctaagtattt gtggccttta
     4261 tcttctacgt agtgaggatc tctcagcgta tggttgtcgc ctgagctgta gttgccttca
     4321 tcgatgaact gctgtacatt ttgatacgtt tttccgtcac cgtcaaagat tgatttataa
     4381 tcctctacac cgttgatgtt caaagagctg tctgatgctg atacgttaac ttgtgcagtt
     4441 gtcagtgttt gtttgccgta atgtttaccg gagaaatcag tgtagaataa acggattttt
     4501 ccgtcagatg taaatgtggc tgaacctgac cattcttgtg tttggtcttt taggatagaa
     4561 tcatttgcat cgaatttgtc gctgtcttta aagacgcggc cagcgttttt ccagctgtca
     4621 atagaagttt cgccgacttt ttgatagaac atgtaaatcg atgtgtcatc cgcattttta
     4681 ggatctccgg ctaatgcaaa gacgatgtgg tagccgtgat agtttgcgac agtgccgtca
     4741 gcgttttgta atggccagct gtcccaaacg tccaggcctt ttgcagaaga gatattttta
     4801 attgtggacg aatcaaattc agaaacttga tatttttcat ttttttgctg ttcagggatt
     4861 tgcagcatat catggcgtgt aatatgggaa atgccgtatg tttccttata tggcttttgg
     4921 ttcgtttctt ccgcaaacgc ttgagttgcg ccgtcgctct gcgcgtgcgc cgacatgcaa
     4981 gcggctgcgg tgagcgcggc ggcgctcagg gccaaacgtt tgtgtctacg ggaaacggct
     5041 tgtttggagt acgtcatcgt ttcctcctaa cgcttaattg ttatccgctc acaattccac
     5101 acattaaact agattcgatg attaattgtc aacagctcac tagtttaatt aatcagaaga
     5161 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     5221 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     5281 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     5341 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     5401 cctcgccgtc gggcatccgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     5461 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     5521 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     5581 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     5641 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     5701 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     5761 cgtcttggag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     5821 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     5881 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     5941 caatcatact cttccttttt caaattattg aagcatttat cagggttatt gtctcatgag
     6001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6061 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6121 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6181 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6241 agcccgtcgc atgcatctag agggatccgc tgcagggcgg ccgctcaggt cagcacggtc
     6301 atgaatcgtt cgttgagaat gcggtcgtgg taaaagatcg ccttgcccag ctggtcggtg
     6361 gtccaggtca ccggtttgtg gtccgggtag ttgtaatctc ccccgcagaa cacttcgttg
     6421 cggttaccgg acgggtcgaa gaagtagatg gtcttgccgt gagtgaggcc gtggcgggtt
     6481 gggccgatat cgatagatgt gtcggtcatg gagatcaggt cggcggcgcg aagcaagtct
     6541 tcccaggttt cgaggtggaa ggacacatga tggaggcggc ctttttccgg atggtgaatg
     6601 aaggccacgt cgtgggcctt ggtcgacaga ctgagaaact gggcgacgcg cgtgccattt
     6661 tcgtccagca cctgttcggc cagatagaaa ccgagcacct tggtgaacag gtcataggtc
     6721 gccggcaatt cgtcgccata catgagggcg tggtcgaaac gcacagccgc catacctttc
     6781 agatcgcgcg gccatgcctc gggattgacg tcattcaaac cccactttcc agtatattcc
     6841 ttgtctgcat acaactcgaa gtgatgcccg gagggggcct ggaagcgcac gcgccggcca
     6901 caactgttca gttcacctgc gggtagctgc tcaacggcac agccatatgc catcagatcc
     6961 cgctccagtt gccggagagc atcctcatcc acaaccttga aacccataaa atccatgccc
     7021 ggctcgtcag cctcgcgtag caccagggaa aacttatcca cttcggtcca agccttcaga
     7081 tagacacggc cctggtcgtc acggtccatc tcgatcaggc ccagcaactc gacgtagtgt
     7141 tccagggcct tgctcatgtc cagtacacgc agctgcacat ggcccggtcg cattacacct
     7201 ttgttcatag tattctgtct ccttctgtga tattccacac attatacgag ccgatgatta
     7261 atggtcaaca gctcaagctt cccgggtacc gagctcgaat tagcttcaaa agcgctctga
     7321 agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga tcccctgatt
     7381 ccctttgtca acagcaatgg atatcagccg ttgacttagt tggtatttcc ggaatatcat
     7441 gctgggttcc gaataatttt gtttaacttt aagaaggaga tataccatgg aagaacaagc
     7501 gtgaacatgg cgcgattgaa gggggaggtt ttgcccgccc tcttcgcgtc gccggcgacg
     7561 attggcgagt acggggccgg catcgacggg gcggattccc tcaacgagct gtcgaatctg
     7621 atggagcacg gcgcagttgc cgcgctggcc gacaaaatca gccagatcgt ggcgaagctg
     7681 gtcgacgcgg acccccgcaa gatcgcggaa aagcctacct ggttcgagaa gatgcttggc
     7741 cgtgaggttg aacgccaggt gaggtatcag gtcgcccgca agacgctcga ccagttgctg
     7801 gacgaagccg agggcgtagc gcagcgcgtg cgggacacgt tgcgcgcctt ggatgacatg
     7861 ctcaatacgc atgaggccga ggtagaccgg ctcagagcct acattcaagc cgggcgcgag
     7921 ttcctggacg agaaccccga ggccggcgcg gccaaggccg gcgtgatcga gttcgacaag
     7981 ccgcgcgaac gcttcgcgcg caagctcgcc aacctggcaa ccctcatggc gtcccatgaa
     8041 atgagcgtca ctcagatgaa gctcacgcgg gcgcaggccg tggacatgct ggaccgcttc
     8101 tctgaaacgg catccgtcct ggtgcccgtc tggcgtcagc acaccctcgc gctcatcacc
     8161 accaagaaca tgaatccggc aatggtcgcc gaggcggcca aagctcacca ggcgctcatg
     8221 cggagccttt cgcagagcct ggaaggcatc aaccaataac acggcgggag aaccctatga
     8281 acgcactgaa aacgacgcac gacgccaagg cccctatcgt cgcctttgac atgaccccgg
     8341 caaccctgcg cgagctgggc ttgcaggaaa gcgacgtgcc ggaagtccat gcggtcgcgc
     8401 agcggatcga ggtcggcagt ccgcagaccg ttgccgagtt cggccgcgac gtggccgagc
     8461 acacgtcccg ctacgccgat agcctgctgg accaggtgcg caacagcgac ctggacgaag
     8521 caggcgagaa actgacccag gttgtcgcca aggcccgttc cctgaacgtc ggccctttgt
     8581 ccgacaaccg ttcccgcctg cccctgattg gcccgctgat cgaccgctcc cgcgtccgtt
     8641 cgacgggctt catggcgcgc ttcgacacga cccgcgagca gatcgaacac ctggtcagcg
     8701 aagtgcagac cacccagcaa ggcatcgcgc agcgcaatgc ctcgctcgac gaaatgttcg
     8761 cagccgtgcg cgaggaacac cgccttcttg gcgtccacat cgcggccggc aaggtccgcc
     8821 ttgccgagct gcgcgagcag gccgagggtc tgcgcggcaa tgtcgggaac gaccctggcc
     8881 gcgtgcagga gctggccgac ctcgatgcga tggttgccaa cctggacaag cgcatcggcg
     8941 acctgatcgc cttgcaacat tcggccatgc agagcctgcc gaccatccgc atgatccagg
     9001 ccaacaacca gatgctggtc gataaattcc acaccatccg cgaaatcacc gtgccggcgt
     9061 ggaagcggca attcatgctg gccttgagcc tcaacgagca gaagaacgcc gtcgaactgg
     9121 ccacggccat cgacgacacc accaacgacc tgatgaagcg caatgcggcc ctgctgcatc
     9181 gcacgtccgt cgagacggcg aaggagaacc aacgcctggt gatcgacgtg gacacgctca
     9241 agcaggttca gacgacgctc atcaagaccg tcgaggacgt tattcgcatc cagcaggaag
     9301 gcgtgcagaa gcgcaaggat gccgagaagc agatcgccgc aatgcgtggc gatcttcaag
     9361 ccaagctgac ccgccagccc gtgcgcgagc tggcccaaca ggagtccgt
//
