LOCUS       V004506                 5461 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004506
VERSION     V004506
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5461)
  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 5461)
  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 5461)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5461)
  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..5461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(352..461)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     terminator      complement(882..909)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1001..1087)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1437..1453
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(1457..1513)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1523..1539)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1547..1563)
                     /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(1571..1601)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1616..1637)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(1688..1706)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1914..2302
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             2455..3450
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     CDS             complement(3862..4662)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(4663..4883)
                     /label="URA3 promoter"
     misc_feature    4911..5414
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgagctaga gtcgatcttc gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg
       61 tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc aggtcgccat
      121 tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt ttgtagccct
      181 ggccgacggc cagcaggtag gccgacaggc tcatgccggc cgccgccgcc ttttcctcaa
      241 tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc ttcctggttg
      301 gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc
      361 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
      421 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
      481 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
      541 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaagcgct
      601 gcttccctgc tgttttgtgg aatatctacc gactggaaac aggcaaatgc aggaaattac
      661 tgaactgagg ggacaggcga gagacgatgc caaagagcta caccgacgag ctggccgagt
      721 gggttgaatc ccgcgcggcc aagaagcgcc ggcgtgatga ggctgcggtt gcgttcctgg
      781 cggtgagggc ggatgtcgat atgcgtaagg agaaaatacc gcatcaggcg catatttgaa
      841 tgtatttaga aaaataaaca aaaagagttt gtagaaacgc aaaaaggcca tccgtcagga
      901 tggccttctg cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
      961 cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     1021 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     1081 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
     1141 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
     1201 ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg
     1261 aaaatcttct ctcatccgcc aaaacagcca agctcgccat tcgccattca ggctgcgcaa
     1321 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     1381 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     1441 aacgacggcc agtgccaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac
     1501 cgagctcgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaaccgct
     1681 ctagacccct atagtgagtc gtattacaaa ctggaacaac actcaaccct atctcggtct
     1741 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1801 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1861 ttttcgggga aatgtgcgcg gaaccagatc cccacgatgc gtccggcgta gaggatctga
     1921 agatcagcag ttcaacctgt tgatagtacg tactaagctc tcatgtttca cgtactaagc
     1981 tctcatgttt aacgtactaa gctctcatgt ttaacgaact aaaccctcat ggctaacgta
     2041 ctaagctctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     2101 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
     2161 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
     2221 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttcagtgaca
     2281 caggaacact taacggctga caggacgaaa gggcctcgtg atacgcgagt tgaaaatttt
     2341 ttttttgact caatatctag acttgcaaga gcttggaact ttgagattgt tctaagatgc
     2401 atacaaattc aaaatgctta aacaaaaata atttaacaaa ggagtaagat tcatatgaca
     2461 caagaatccc tgttacttct cgaccgtatt gattcggatg attcctacgc gagcctgcgg
     2521 aacgaccagg aattctggga gccgctggcc cgccgagccc tggaggagct cgggctgccg
     2581 gtgccgccgg tgctgcgggt gcccggcgag agcaccaacc ccgtactggt cggcgagccc
     2641 gacccggtga tcaagctgtt cggcgagcac tggtgcggtc cggagagcct cgcgtcggag
     2701 tcggaggcgt acgcggtcct ggcggacgcc ccggtgccgg tgccccgcct cctcggccgc
     2761 ggcgagctgc ggcccggcac cggagcctgg ccgtggccct acctggtgat gagccggatg
     2821 accggcacca cctggcggtc cgcgatggac ggcacgaccg accggaacgc gctgctcgcc
     2881 ctggcccgcg aactcggccg ggtgctcggc cggctgcaca gggtgccgct gaccgggaac
     2941 accgtgctca ccccccattc cgaggtcttc ccggaactgc tgcgggaacg ccgcgcggcg
     3001 accgtcgagg accaccgcgg gtggggctac ctctcgcccc ggctgctgga ccgcctggag
     3061 gactggctgc cggacgtgga cacgctgctg gccggccgcg aaccccggtt cgtccacggc
     3121 gacctgcacg ggaccaacat cttcgtggac ctggccgcga ccgaggtcac cgggatcgtc
     3181 gacttcaccg acgtctatgc gggagactcc cgctacagcc tggtgcaact gcatctcaac
     3241 gccttccggg gcgaccgcga gatcctggcc gcgctgctcg acggggcgca gtggaagcgg
     3301 accgaggact tcgcccgcga actgctcgcc ttcaccttcc tgcacgactt cgaggtgttc
     3361 gaggagaccc cgctggatct ctccggcttc accgatccgg aggaactggc gcagttcctc
     3421 tgggggccgc cggacaccgc ccccggcgcc tgaacgtaac tgtcagacca agtttactca
     3481 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3541 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3601 gaccccgtag aaaagatcaa aggatcttct tgagatccag ttacgctagg gataacaggg
     3661 taatatagct cgagattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3781 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     3841 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     3901 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     3961 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     4021 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     4081 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     4141 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     4201 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     4261 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     4321 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     4381 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     4441 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     4501 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     4561 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     4621 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct
     4681 gcaggttttt gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac
     4741 actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     4801 ttcggagatt accgaatcaa aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa
     4861 aaaaaatgat gaattgaatt gaaaagctag cttatcgatg ggtccttttc atcacgtgct
     4921 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     4981 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     5041 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     5101 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac
     5161 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     5221 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     5281 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     5341 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     5401 acaggcaagc gatctctaag aaaccattat tatcatgaca ttaacctata aaaaaggcct
     5461 c
//
