LOCUS       V004501                 8422 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004501
VERSION     V004501
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8422)
  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 8422)
  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 8422)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8422)
  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..8422
                     /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"
     CDS             complement(1170..1187)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             complement(1197..1238)
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             complement(1269..4313)
                     /label="lacZ"
                     /note="beta-galactosidase"
     protein_bind    complement(4361..4377)
                     /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(4385..4415)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4430..4451)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(4502..4520)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      4728..5116
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             5559..6554
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     CDS             complement(6823..7623)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(7624..7844)
                     /label="URA3 promoter"
     misc_feature    7872..8375
                     /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 ctcctctagg atcagcgggt ttaaactcaa tggtgatggt gatgatgacc ggtacgcgta
     1201 gaatcgagac cgaggagagg gttagggata ggcttacctt cgaagggccc tctagactcg
     1261 agcggccgct tttgacacca gaccaactgg taatggtagc gaccggcgct cagctggaat
     1321 tccgccgata ctgacgggct ccaggagtcg tcgccaccaa tccccatatg gaaaccgtcg
     1381 atattcagcc atgtgccttc ttccgcgtgc agcagatggc gatggctggt ttccatcagt
     1441 tgctgttgac tgtagcggct gatgttgaac tggaagtcgc cgcgccactg gtgtgggcca
     1501 taattcaatt cgcgcgtccc gcagcgcaga ccgttttcgc tcgggaagac gtacggggta
     1561 tacatgtctg acaatggcag atcccagcgg tcaaaacagg cggcagtaag gcggtcggga
     1621 tagttttctt gcggccctaa tccgagccag tttacccgct ctgctacctg cgccagctgg
     1681 cagttcaggc caatccgcgc cggatgcggt gtatcgctcg ccacttcaac atcaacggta
     1741 atcgccattt gaccactacc atcaatccgg taggttttcc ggctgataaa taaggttttc
     1801 ccctgatgct gccacgcgtg agcggtcgta atcagcaccg catcagcaag tgtatctgcc
     1861 gtgcactgca acaacgctgc ttcggcctgg taatggcccg ccgccttcca gcgttcgacc
     1921 caggcgttag ggtcaatgcg ggtcgcttca cttacgccaa tgtcgttatc cagcggtgca
     1981 cgggtgaact gatcgcgcag cggcgtcagc agttgttttt tatcgccaat ccacatctgt
     2041 gaaagaaagc ctgactggcg gttaaattgc caacgcttat tacccagctc gatgcaaaaa
     2101 tccatttcgc tggtggtcag atgcgggatg gcgtgggacg cggcggggag cgtcacactg
     2161 aggttttccg ccagacgcca ctgctgccag gcgctgatgt gcccggcttc tgaccatgcg
     2221 gtcgcgttcg gttgcactac gcgtactgtg agccagagtt gcccggcgct ctccggctgc
     2281 ggtagttcag gcagttcaat caactgttta ccttgtggag cgacatccag aggcacttca
     2341 ccgcttgcca gcggcttacc atccagcgcc accatccagt gcaggagctc gttatcgcta
     2401 tgacggaaca ggtattcgct ggtcacttcg atggtttgcc cggataaacg gaactggaaa
     2461 aactgctgct ggtgttttgc ttccgtcagc gctggatgcg gcgtgcggtc ggcaaagacc
     2521 agaccgttca tacagaactg gcgatcgttc ggcgtatcgc caaaatcacc gccgtaagcc
     2581 gaccacgggt tgccgttttc atcatattta atcagcgact gatccaccca gtcccagacg
     2641 aagccgccct gtaaacgggg atactgacga aacgcctgcc agtatttagc gaaaccgcca
     2701 agactgttac ccatcgcgtg ggcgtattcg caaaggatca gcgggcgcgt ctctccaggt
     2761 agcgaaagcc attttttgat ggaccatttc ggcacagccg ggaagggctg gtcttcatcc
     2821 acgcgcgcgt acatcgggca aataatatcg gtggccgtgg tgtcggctcc gccgccttca
     2881 tactgcaccg ggcgggaagg atcgacagat ttgatccagc gatacagcgc gtcgtgatta
     2941 gcgccgtggc ctgattcatt ccccagcgac cagatgatca cactcgggtg attacgatcg
     3001 cgctgcacca ttcgcgttac gcgttcgctc atcgccggta gccagcgcgg atcatcggtc
     3061 agacgattca ttggcaccat gccgtgggtt tcaatattgg cttcatccac cacatacagg
     3121 ccgtagcggt cgcacagcgt gtaccacagc ggatggttcg gataatgcga acagcgcacg
     3181 gcgttaaagt tgttctgctt catcagcagg atatcctgca ccatcgtctg ctcatccatg
     3241 acctgaccat gcagaggatg atgctcgtga cggttaacgc ctcgaatcag caacggcttg
     3301 ccgttcagca gcagcagacc attttcaatc cgcacctcgc ggaaaccgac atcgcaggct
     3361 tctgcttcaa tcagcgtgcc gtcggcggtg tgcagttcaa ccaccgcacg atagagattc
     3421 gggatttcgg cgctccacag tttcgggttt tcgacgttca gacgtagtgt gacgcgatcg
     3481 gcataaccac cacgctcatc gataatttca ccgccgaaag gcgcggtgcc gctggcgacc
     3541 tgcgtttcac cctgccataa agaaactgtt acccgtaggt agtcacgcaa ctcgccgcac
     3601 atctgaactt cagcctccag tacagcgcgg ctgaaatcat cattaaagcg agtggcaaca
     3661 tggaaatcgc tgatttgtgt agtcggttta tgcagcaacg agacgtcacg gaaaatgccg
     3721 ctcatccgcc acatatcctg atcttccaga taactgccgt cactccaacg cagcaccatc
     3781 accgcgaggc ggttttctcc ggcgcgtaaa aatgcgctca ggtcaaattc agacggcaaa
     3841 cgactgtcct ggccgtaacc gacccagcgc ccgttgcacc acagatgaaa cgccgagtta
     3901 acgccatcaa aaataattcg cgtctggcct tcctgtagcc agctttcatc aacattaaat
     3961 gtgagcgagt aacaacccgt cggattctcc gtgggaacaa acggcggatt gaccgtaatg
     4021 ggataggtta cgttggtgta gatgggcgca tcgtaaccgt gcatctgcca gtttgagggg
     4081 acgacgacag tatcggcctc aggaagatcg cactccagcc agctttccgg caccgcttct
     4141 ggtgccggaa accaggcaaa gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4201 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     4261 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacgggatct
     4321 atcatcgtgg atccgagctc ggtacctcct ctgtgtgaaa ttgttatccg ctcacaattc
     4381 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4441 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac cgctctagac
     4501 ccctatagtg agtcgtatta caaactggaa caacactcaa ccctatctcg gtctattctt
     4561 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4621 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4681 gggaaatgtg cgcggaacca gatccccacg atgcgtccgg cgtagaggat ctgaagatca
     4741 gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact aagctctcat
     4801 gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc
     4861 tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa
     4921 taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa
     4981 aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag
     5041 ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttcagt gacacaggaa
     5101 cacttaacgg ctgacagacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
     5161 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     5221 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     5281 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     5341 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     5401 ccgccaagga tctgatggcg caggggatcc gagttgaaaa tttttttttt gactcaatat
     5461 ctagacttgc aagagcttgg aactttgaga ttgttctaag atgcatacaa attcaaaatg
     5521 cttaaacaaa aataatttaa caaaggagta agattcatat gacacaagaa tccctgttac
     5581 ttctcgaccg tattgattcg gatgattcct acgcgagcct gcggaacgac caggaattct
     5641 gggagccgct ggcccgccga gccctggagg agctcgggct gccggtgccg ccggtgctgc
     5701 gggtgcccgg cgagagcacc aaccccgtac tggtcggcga gcccgacccg gtgatcaagc
     5761 tgttcggcga gcactggtgc ggtccggaga gcctcgcgtc ggagtcggag gcgtacgcgg
     5821 tcctggcgga cgccccggtg ccggtgcccc gcctcctcgg ccgcggcgag ctgcggcccg
     5881 gcaccggagc ctggccgtgg ccctacctgg tgatgagccg gatgaccggc accacctggc
     5941 ggtccgcgat ggacggcacg accgaccgga acgcgctgct cgccctggcc cgcgaactcg
     6001 gccgggtgct cggccggctg cacagggtgc cgctgaccgg gaacaccgtg ctcacccccc
     6061 attccgaggt cttcccggaa ctgctgcggg aacgccgcgc ggcgaccgtc gaggaccacc
     6121 gcgggtgggg ctacctctcg ccccggctgc tggaccgcct ggaggactgg ctgccggacg
     6181 tggacacgct gctggccggc cgcgaacccc ggttcgtcca cggcgacctg cacgggacca
     6241 acatcttcgt ggacctggcc gcgaccgagg tcaccgggat cgtcgacttc accgacgtct
     6301 atgcgggaga ctcccgctac agcctggtgc aactgcatct caacgccttc cggggcgacc
     6361 gcgagatcct ggccgcgctg ctcgacgggg cgcagtggaa gcggaccgag gacttcgccc
     6421 gcgaactgct cgccttcacc ttcctgcacg acttcgaggt gttcgaggag accccgctgg
     6481 atctctccgg cttcaccgat ccggaggaac tggcgcagtt cctctggggg ccgccggaca
     6541 ccgcccccgg cgcctgaacg tggttctccg ttaagtaagg ccaaacgttt taacttaaat
     6601 gtcaaactaa actcgtagag ttttggacaa tcctgattat tgaagcattt atcagggtta
     6661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6721 gcgcacattt ccccgaaaag tgggtaataa ctgatataat taaattgaag ctctaatttg
     6781 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     6841 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     6901 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     6961 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     7021 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     7081 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     7141 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     7201 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     7261 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     7321 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     7381 ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac
     7441 atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt
     7501 ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa
     7561 tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga
     7621 catgatttat cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg
     7681 gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc
     7741 cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg
     7801 aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagcta gcttatcgat
     7861 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat
     7921 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga
     7981 agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg
     8041 ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa
     8101 tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt
     8161 cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt
     8221 tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc taaaatccaa
     8281 atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc atcattaaaa
     8341 gatacgaggc gcgtgtaagt tacaggcaag cgatctctaa gaaaccatta ttatcatgac
     8401 attaacctat aaaaaaggcc tc
//
