LOCUS       V004507                 6432 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004507
VERSION     V004507
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6432)
  AUTHORS   Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ,
            Shanks RM.
  TITLE     New vector tools with a hygromycin resistance marker for use with
            opportunistic pathogens
  JOURNAL   Mol. Biotechnol. 48 (1), 7-14 (2011)
   PUBMED   20972648
REFERENCE   2  (bases 1 to 6432)
  AUTHORS   Kalivoda EJ, Horzempa JE, Sadaf AA, Stella NA, Nau GJ, Shanks RM.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI
            1015, 203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE   3  (bases 1 to 6432)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6432)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
            Biotechnol."; date: "2011"; volume: "48"; issue: "1"; pages: "7-14"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI 1015, 203
            Lothrop St, Pittsburgh, PA 15213, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(399..508)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     terminator      complement(929..956)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1048..1134)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1484..1500
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(1504..1560)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1570..1586)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1594..1610)
                     /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)."
     promoter        complement(1618..1648)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1663..1684)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      1834..2603
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     CDS             2604..3263
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     CDS             complement(3550..4545)
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(4676..4780)
                     /label="AmpR promoter"
     CDS             complement(4880..5680)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5681..5901)
                     /label="URA3 promoter"
     misc_feature    5929..6432
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa aggcctctcg agctagagtc
       61 gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc
      121 ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag
      181 ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag
      241 caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc
      301 gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc
      361 agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg
      421 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
      481 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
      541 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
      601 aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct tccctgctgt
      661 tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga actgagggga
      721 caggcgagag acgatgccaa agagctacac cgacgagctg gccgagtggg ttgaatcccg
      781 cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg tgagggcgga
      841 tgtcgatatg cgtaaggaga aaataccgca tcaggcgcat atttgaatgt atttagaaaa
      901 ataaacaaaa agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      961 aatttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     1021 cgcaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     1081 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     1141 ggcagttccc tactctcgca tggggagacc ccacactacc atcggcgcta cggcgtttca
     1201 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct
     1261 gttttatcag accgcttctg cgttctgatt taatctgtat caggctgaaa atcttctctc
     1321 atccgccaaa acagccaagc tcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     1381 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     1441 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     1501 gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc
     1561 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1621 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1681 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1741 ttaatgcgcc ctacgggctt gctctccggg cttcgccctg cgcggtcgct gcgctccctt
     1801 gccagcccgt ggatatgtgg acgatggccg cgagcggcca ccggctggct cgcttcgctc
     1861 ggcccgtgga caaccctgct ggacaagctg atggacaggc tgcgcctgcc cacgagcttg
     1921 accacaggga ttgcccaccg gctacccagc cttcgaccac atacccaccg gctccgactg
     1981 cgcggcctgc ggccttgccc catcaatttt tttaattttc tctggggaaa agcctccggc
     2041 ctgcggcctg cgcgcttcgc ttgccggttg gacaccaagt ggaaggcggg tcaaggctcg
     2101 cgcagcgacc gcgcagcggc ttggccttga cgcgcctgga acgacccaag cctatgcgag
     2161 tgggggcagt cgaaggcgaa gcccgcccgc ctgccccccg agcctcacgg cggcgagtgc
     2221 gggggttcca agggggcagc gccaccttgg gcaaggccga aggccgcgca gtcgatcaac
     2281 aagccccgga ggggccactt tttgccggag ggggagccgc gccgaaggcg tgggggaacc
     2341 ccgcaggggt gcccttcttt gggcaccaaa gaactagata tagggcgaaa tgcgaaagac
     2401 ttaaaaatca acaacttaaa aaaggggggt acgcaacagc tcattgcggc accccccgca
     2461 atagctcatt gcgtaggtta aagaaaatct gtaattgact gccactttta cgcaacgcat
     2521 aattgttgtc gcgctgccga aaagttgcag ctgattgcgc atggtgccgc aaccgtgcgg
     2581 caccctaccg catggagata agcatggcca cgcagtccag agaaatcggc attcaagcca
     2641 agaacaagcc cggtcactgg gtgcaaacgg aacgcaaagc gcatgaggcg tgggccgggc
     2701 ttattgcgag gaaacccacg gcggcaatgc tgctgcatca cctcgtggcg cagatgggcc
     2761 accagaacgc cgtggtggtc agccagaaga cactttccaa gctcatcgga cgttctttgc
     2821 ggacggtcca atacgcagtc aaggacttgg tggccgagcg ctggatctcc gtcgtgaagc
     2881 tcaacggccc cggcaccgtg tcggcctacg tggtcaatga ccgcgtggcg tggggccagc
     2941 cccgcgacca gttgcgcctg tcggtgttca gtgccgccgt ggtggttgat cacgacgacc
     3001 aggacgaatc gctgttgggg catggcgacc tgcgccgcat cccgaccctg tatccgggcg
     3061 agcagcaact accgaccggc cccggcgagg agccgcccag ccagcccggc attccgggca
     3121 tggaaccaga cctgccagcc ttgaccgaaa cggaggaatg ggaacggcgc gggcagcagc
     3181 gcctgccgat gcccgatgag ccgtgttttc tggacgatgg cgagccgttg gagccgccga
     3241 cacgggtcac gctgccgcgc cggtagcact tgggttgcgc agcaacccgt aagtgcgctg
     3301 ttccagacta tcggctgtag ccgcctcgcc gccctatacc ttgtctgcct ccccgcgttg
     3361 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagcctttta cgagtttttg
     3421 gtgcaatcaa tctaaagtat atatgagtaa acttggtctg acaatcgatg cgaattggcc
     3481 gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac
     3541 gaattgtcag gcgccggggg cggtgtccgg cggcccccag aggaactgcg ccagttcctc
     3601 cggatcggtg aagccggaga gatccagcgg ggtctcctcg aacacctcga agtcgtgcag
     3661 gaaggtgaag gcgagcagtt cgcgggcgaa gtcctcggtc cgcttccact gcgccccgtc
     3721 gagcagcgcg gccaggatct cgcggtcgcc ccggaaggcg ttgagatgca gttgcaccag
     3781 gctgtagcgg gagtctcccg catagacgtc ggtgaagtcg acgatcccgg tgacctcggt
     3841 cgcggccagg tccacgaaga tgttggtccc gtgcaggtcg ccgtggacga accggggttc
     3901 gcggccggcc agcagcgtgt ccacgtccgg cagccagtcc tccaggcggt ccagcagccg
     3961 gggcgagagg tagccccacc cgcggtggtc ctcgacggtc gccgcgcggc gttcccgcag
     4021 cagttccggg aagacctcgg aatggggggt gagcacggtg ttcccggtca gcggcaccct
     4081 gtgcagccgg ccgagcaccc ggccgagttc gcgggccagg gcgagcagcg cgttccggtc
     4141 ggtcgtgccg tccatcgcgg accgccaggt ggtgccggtc atccggctca tcaccaggta
     4201 gggccacggc caggctccgg tgccgggccg cagctcgccg cggccgagga ggcggggcac
     4261 cggcaccggg gcgtccgcca ggaccgcgta cgcctccgac tccgacgcga ggctctccgg
     4321 accgcaccag tgctcgccga acagcttgat caccgggtcg ggctcgccga ccagtacggg
     4381 gttggtgctc tcgccgggca cccgcagcac cggcggcacc ggcagcccga gctcctccag
     4441 ggctcggcgg gccagcggct cccagaattc ctggtcgttc cgcaggctcg cgtaggaatc
     4501 atccgaatca atacggtcga gaagtaacag ggattcttgt gtcatatgaa tcttactcct
     4561 ttgttaaatt atttttgttt aagcattttg aatttgtatg catcttagaa caatctcaaa
     4621 gttccaagct cttgcaagtc tagatattga gtcaaaaaaa aaattttcaa ctcatactct
     4681 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4741 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgg
     4801 gtaataactg atataattaa attgaagctc taatttgtga gtttagtata catgcattta
     4861 cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg
     4921 cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc
     4981 caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac
     5041 ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac
     5101 cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc
     5161 tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg
     5221 acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct
     5281 tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt
     5341 ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa
     5401 attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa
     5461 ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt
     5521 tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag
     5581 cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag
     5641 gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc
     5701 aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac
     5761 tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt
     5821 cggagattac cgaatcaaaa aaatttcaaa gaaaccgaaa tcaaaaaaaa gaataaaaaa
     5881 aaaatgatga attgaattga aaagctagct tatcgatggg tccttttcat cacgtgctat
     5941 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     6001 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     6061 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     6121 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt
     6181 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     6241 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     6301 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     6361 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     6421 aggcaagcga tc
//
