LOCUS       V004510                 6261 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004510
VERSION     V004510
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6261)
  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 6261)
  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 6261)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6261)
  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..6261
                     /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      1871..2093
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2141..3088
                     /label="Rep101(Ts)"
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
     CDS             complement(3379..4374)
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(4505..4609)
                     /label="AmpR promoter"
     CDS             complement(4709..5509)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5510..5730)
                     /label="URA3 promoter"
     misc_feature    5758..6261
                     /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 ttaatgcagc gatttgcccg agcttgcgag ggtgctactt aagcctttag ggttttaagg
     1801 tctgttttgt agaggagcaa acagcgtttg cgacatcctt ttgtaatact gcggaactga
     1861 ctaaagtagt gagttataca cagggctggg atctattctt tttatctttt tttattcttt
     1921 ctttattcta taaattataa ccacttgaat ataaacaaaa aaaacacaca aaggtctagc
     1981 ggaatttaca gagggtctag cagaatttac aagttttcca gcaaaggtct agcagaattt
     2041 acagataccc acaactcaaa ggaaaaggac tagtaattat cattgactag cccatctcaa
     2101 ttggtatagt gattaaaatc acctagacca attgagatgt atgtctgaat tagttgtttt
     2161 caaagcaaat gaactagcga ctagtcgcta tgacttaacg gagcatgaaa ccaagctaat
     2221 tttatgctgt gtggcactac tcaaccccac gattgaaaac cctacaagga aagaacggac
     2281 ggtatcgttc acttataacc aatacgttca gatgatgaac atcagtaggg aaaatgctta
     2341 tggtgtatta gctaaagcaa ccagagagct gatgacgaga actgtggaaa tcaggaatcc
     2401 tttggttaaa ggctttgaga ttttccagtg gacaaactat gccaagttct caagcgaaaa
     2461 attagaatta gtttttagtg aagagatatt gccttatctt ttccagttaa aaaaattcat
     2521 aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa tactctatga ggatttatga
     2581 gtggttatta aaagaactaa cacaaaagaa aactcacaag gcaaatatag agattagcct
     2641 tgatgaattt aagttcatgt taatgcttga aaataactac catgagttta aaaggcttaa
     2701 ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact tacagcaata tgaaattggt
     2761 ggttgataag cgaggccgcc cgactgatac gttgattttc caagttgaac tagatagaca
     2821 aatggatctc gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg acaaaatacc
     2881 aacaaccatt acatcagatt cctacctaca taacggacta agaaaaacac tacacgatgc
     2941 tttaactgca aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag
     3001 taagtatgat ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa ccacactaga
     3061 gaacatactg gctaaatacg gaaggatctg aggttcttat ggctcttgta tctatcagtg
     3121 aagcatcaag actaacaaac aaaagtagaa caactgttca ccgttacata tcaaagggaa
     3181 aactgtccat atgcacagat gaaaacggtg taaaaaagat agatacatca gagcttttac
     3241 gagtttttgg tgcaatcaat ctaaagtata tatgagtaaa cttggtctga caatcgatgc
     3301 gaattggccg cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc
     3361 ggcttgaacg aattgtcagg cgccgggggc ggtgtccggc ggcccccaga ggaactgcgc
     3421 cagttcctcc ggatcggtga agccggagag atccagcggg gtctcctcga acacctcgaa
     3481 gtcgtgcagg aaggtgaagg cgagcagttc gcgggcgaag tcctcggtcc gcttccactg
     3541 cgccccgtcg agcagcgcgg ccaggatctc gcggtcgccc cggaaggcgt tgagatgcag
     3601 ttgcaccagg ctgtagcggg agtctcccgc atagacgtcg gtgaagtcga cgatcccggt
     3661 gacctcggtc gcggccaggt ccacgaagat gttggtcccg tgcaggtcgc cgtggacgaa
     3721 ccggggttcg cggccggcca gcagcgtgtc cacgtccggc agccagtcct ccaggcggtc
     3781 cagcagccgg ggcgagaggt agccccaccc gcggtggtcc tcgacggtcg ccgcgcggcg
     3841 ttcccgcagc agttccggga agacctcgga atggggggtg agcacggtgt tcccggtcag
     3901 cggcaccctg tgcagccggc cgagcacccg gccgagttcg cgggccaggg cgagcagcgc
     3961 gttccggtcg gtcgtgccgt ccatcgcgga ccgccaggtg gtgccggtca tccggctcat
     4021 caccaggtag ggccacggcc aggctccggt gccgggccgc agctcgccgc ggccgaggag
     4081 gcggggcacc ggcaccgggg cgtccgccag gaccgcgtac gcctccgact ccgacgcgag
     4141 gctctccgga ccgcaccagt gctcgccgaa cagcttgatc accgggtcgg gctcgccgac
     4201 cagtacgggg ttggtgctct cgccgggcac ccgcagcacc ggcggcaccg gcagcccgag
     4261 ctcctccagg gctcggcggg ccagcggctc ccagaattcc tggtcgttcc gcaggctcgc
     4321 gtaggaatca tccgaatcaa tacggtcgag aagtaacagg gattcttgtg tcatatgaat
     4381 cttactcctt tgttaaatta tttttgttta agcattttga atttgtatgc atcttagaac
     4441 aatctcaaag ttccaagctc ttgcaagtct agatattgag tcaaaaaaaa aattttcaac
     4501 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4561 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4621 gaaaagtggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     4681 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     4741 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     4801 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     4861 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     4921 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     4981 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     5041 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     5101 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     5161 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     5221 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     5281 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     5341 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     5401 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     5461 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     5521 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     5581 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     5641 ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag
     5701 aataaaaaaa aaatgatgaa ttgaattgaa aagctagctt atcgatgggt ccttttcatc
     5761 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     5821 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     5881 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     5941 tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac
     6001 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     6061 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     6121 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     6181 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     6241 gtaagttaca ggcaagcgat c
//
