LOCUS       V004505                 7084 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004505
VERSION     V004505
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7084)
  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 7084)
  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 7084)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7084)
  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..7084
                     /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"
     CDS             complement(1524..2216)
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     protein_bind    complement(2246..2262)
                     /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(2270..2300)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2315..2336)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      2486..3255
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     CDS             3256..3915
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     CDS             complement(4202..5197)
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(5328..5432)
                     /label="AmpR promoter"
     CDS             complement(5532..6332)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(6333..6553)
                     /label="URA3 promoter"
     misc_feature    6581..7084
                     /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 catgccctta ttatttcagg aaagtttcgg aggagatagt gttcggcagt
     1561 ttgtacatca tctgcgggat caggtacggt ttgatcaggt tgtagaagat caggtaagac
     1621 atagaatcga tgtagatgat cggtttgttt ttgttgattt ttacgtaaca gttcagttgg
     1681 aatttgttac gcagaccctt aaccaggtat tctacttctt cgaaagtgaa agactgggtg
     1741 ttcagtacga tcgatttgtt ggtagagttt ttgttgtaat cccatttacc accatcatcc
     1801 atgaaccagt atgccagaga catcggggtc aggtagtttt caaccaggtt gttcgggatg
     1861 gtttttttgt tgttaacgat gaacaggtta gccagtttgt tgaaagcttg gtgtttgaaa
     1921 gtctgggcgc cccaggtgat taccaggtta cccaggtggt taacacgttc ttttttgtgc
     1981 ggcggggaca gtacccactg atcgtacagc agacatacgt ggtccatgta tgctttgttt
     2041 ttccactcga actgcataca gtaggtttta ccttcatcac gagaacggat gtaagcatca
     2101 cccaggatca gaccgatacc tgcttcgaac tgttcgatgt tcagttcgat cagctgggat
     2161 ttgtattctt tcagcagttt agagttcgga cccaggttca ttacctggtt tttttgatgc
     2221 atatgcgttc tcctcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2281 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2341 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgcg
     2401 ccctacgggc ttgctctccg ggcttcgccc tgcgcggtcg ctgcgctccc ttgccagccc
     2461 gtggatatgt ggacgatggc cgcgagcggc caccggctgg ctcgcttcgc tcggcccgtg
     2521 gacaaccctg ctggacaagc tgatggacag gctgcgcctg cccacgagct tgaccacagg
     2581 gattgcccac cggctaccca gccttcgacc acatacccac cggctccgac tgcgcggcct
     2641 gcggccttgc cccatcaatt tttttaattt tctctgggga aaagcctccg gcctgcggcc
     2701 tgcgcgcttc gcttgccggt tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga
     2761 ccgcgcagcg gcttggcctt gacgcgcctg gaacgaccca agcctatgcg agtgggggca
     2821 gtcgaaggcg aagcccgccc gcctgccccc cgagcctcac ggcggcgagt gcgggggttc
     2881 caagggggca gcgccacctt gggcaaggcc gaaggccgcg cagtcgatca acaagccccg
     2941 gaggggccac tttttgccgg agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg
     3001 gtgcccttct ttgggcacca aagaactaga tatagggcga aatgcgaaag acttaaaaat
     3061 caacaactta aaaaaggggg gtacgcaaca gctcattgcg gcaccccccg caatagctca
     3121 ttgcgtaggt taaagaaaat ctgtaattga ctgccacttt tacgcaacgc ataattgttg
     3181 tcgcgctgcc gaaaagttgc agctgattgc gcatggtgcc gcaaccgtgc ggcaccctac
     3241 cgcatggaga taagcatggc cacgcagtcc agagaaatcg gcattcaagc caagaacaag
     3301 cccggtcact gggtgcaaac ggaacgcaaa gcgcatgagg cgtgggccgg gcttattgcg
     3361 aggaaaccca cggcggcaat gctgctgcat cacctcgtgg cgcagatggg ccaccagaac
     3421 gccgtggtgg tcagccagaa gacactttcc aagctcatcg gacgttcttt gcggacggtc
     3481 caatacgcag tcaaggactt ggtggccgag cgctggatct ccgtcgtgaa gctcaacggc
     3541 cccggcaccg tgtcggccta cgtggtcaat gaccgcgtgg cgtggggcca gccccgcgac
     3601 cagttgcgcc tgtcggtgtt cagtgccgcc gtggtggttg atcacgacga ccaggacgaa
     3661 tcgctgttgg ggcatggcga cctgcgccgc atcccgaccc tgtatccggg cgagcagcaa
     3721 ctaccgaccg gccccggcga ggagccgccc agccagcccg gcattccggg catggaacca
     3781 gacctgccag ccttgaccga aacggaggaa tgggaacggc gcgggcagca gcgcctgccg
     3841 atgcccgatg agccgtgttt tctggacgat ggcgagccgt tggagccgcc gacacgggtc
     3901 acgctgccgc gccggtagca cttgggttgc gcagcaaccc gtaagtgcgc tgttccagac
     3961 tatcggctgt agccgcctcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     4021 tgcatggagc cgggccacct cgacctgaat ggaagccttt tacgagtttt tggtgcaatc
     4081 aatctaaagt atatatgagt aaacttggtc tgacaatcga tgcgaattgg ccgcggcgtt
     4141 gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtc
     4201 aggcgccggg ggcggtgtcc ggcggccccc agaggaactg cgccagttcc tccggatcgg
     4261 tgaagccgga gagatccagc ggggtctcct cgaacacctc gaagtcgtgc aggaaggtga
     4321 aggcgagcag ttcgcgggcg aagtcctcgg tccgcttcca ctgcgccccg tcgagcagcg
     4381 cggccaggat ctcgcggtcg ccccggaagg cgttgagatg cagttgcacc aggctgtagc
     4441 gggagtctcc cgcatagacg tcggtgaagt cgacgatccc ggtgacctcg gtcgcggcca
     4501 ggtccacgaa gatgttggtc ccgtgcaggt cgccgtggac gaaccggggt tcgcggccgg
     4561 ccagcagcgt gtccacgtcc ggcagccagt cctccaggcg gtccagcagc cggggcgaga
     4621 ggtagcccca cccgcggtgg tcctcgacgg tcgccgcgcg gcgttcccgc agcagttccg
     4681 ggaagacctc ggaatggggg gtgagcacgg tgttcccggt cagcggcacc ctgtgcagcc
     4741 ggccgagcac ccggccgagt tcgcgggcca gggcgagcag cgcgttccgg tcggtcgtgc
     4801 cgtccatcgc ggaccgccag gtggtgccgg tcatccggct catcaccagg tagggccacg
     4861 gccaggctcc ggtgccgggc cgcagctcgc cgcggccgag gaggcggggc accggcaccg
     4921 gggcgtccgc caggaccgcg tacgcctccg actccgacgc gaggctctcc ggaccgcacc
     4981 agtgctcgcc gaacagcttg atcaccgggt cgggctcgcc gaccagtacg gggttggtgc
     5041 tctcgccggg cacccgcagc accggcggca ccggcagccc gagctcctcc agggctcggc
     5101 gggccagcgg ctcccagaat tcctggtcgt tccgcaggct cgcgtaggaa tcatccgaat
     5161 caatacggtc gagaagtaac agggattctt gtgtcatatg aatcttactc ctttgttaaa
     5221 ttatttttgt ttaagcattt tgaatttgta tgcatcttag aacaatctca aagttccaag
     5281 ctcttgcaag tctagatatt gagtcaaaaa aaaaattttc aactcatact cttccttttt
     5341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gggtaataac
     5461 tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat
     5521 acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg
     5581 taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata
     5641 ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg
     5701 tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct
     5761 cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca
     5821 atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca tgacaattct
     5881 gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg
     5941 ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt
     6001 ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg
     6061 tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc
     6121 tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct
     6181 aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag
     6241 tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta
     6301 gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct gcaggttttt
     6361 gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg
     6421 cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt
     6481 accgaatcaa aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat
     6541 gaattgaatt gaaaagctag cttatcgatg ggtccttttc atcacgtgct ataaaaataa
     6601 ttataattta aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa
     6661 ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac
     6721 aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc
     6781 ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa
     6841 tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc
     6901 tttttgttga aattttttaa acctttgttt attttttttt cttcattccg taactcttct
     6961 accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag
     7021 taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc
     7081 gatc
//
