LOCUS       V004526                 5980 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004526
VERSION     V004526
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5980)
  AUTHORS   Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
  TITLE     New yeast recombineering tools for bacteria
  JOURNAL   Plasmid 62 (2), 88-97 (2009)
   PUBMED   19477196
REFERENCE   2  (bases 1 to 5980)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
  TITLE     In vivo recombination vectors for modification and expression of
            genes in Gram-negative and Gram-positive bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5980)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAR-2008) Ophthalmology, University of Pittsburgh, 203
            Lothrop St., Pittsburgh, PA 15213, USA
REFERENCE   4  (bases 1 to 5980)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5980)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
            date: "2009"; volume: "62"; issue: "2"; pages: "88-97"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (03-MAR-2008) Ophthalmology, University of Pittsburgh, 203 Lothrop
            St., Pittsburgh, PA 15213, USA"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5980
                     /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             2653..3444
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             3744..4115
                     /gene="rpsL"
                     /label="Small ribosomal subunit protein uS12"
                     /note="Small ribosomal subunit protein uS12 from Salmonella
                     paratyphi A (strain ATCC 9150 / SARB42). Accession#:
                     Q5PIW1"
     CDS             complement(4381..5181)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5182..5402)
                     /label="URA3 promoter"
     misc_feature    5430..5933
                     /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 cagacgctgc cgcaagcact cagggcgcaa gggctgctaa
     2341 aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc
     2401 agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc
     2461 agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa
     2521 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct
     2581 ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga
     2641 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     2701 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     2761 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     2821 ctgaatgaac tccaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     2881 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     2941 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3001 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3061 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3121 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     3181 cggatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     3241 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     3301 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     3361 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     3421 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gtacgcgtct aggcggccgc
     3481 gaagttccta ttccgaagtt cctctagaag cttctgcaga cgggtttgac tggtcaaatt
     3541 tcgaacgaca tcattctcat tgactatact gatttcgtca gacttacggt taagcacccc
     3601 agccagatgg cctggtgatg gcgggatcgt tgtatatttc ttgacacctt ttcggcatcg
     3661 ccctaaaatt cggcgtcctc atattgtgtg aggacgtttt attacgtgtt tacgaagcaa
     3721 aagctaaaac caggagctat ttaatggcaa cagttaacca gctggtacgc aaaccacgtg
     3781 ctcgcaaagt tgcgaaaagc aacgtgcctg cgctggaagc atgcccgcaa aaacgtggcg
     3841 tatgtactcg tgtatatact accactccta aaaaaccgaa ctccgcgctg cgtaaagtat
     3901 gccgtgttcg tctgactaac ggtttcgaag tgacttccta catcggtggt gaaggtcaca
     3961 acctgcagga gcactccgtg atcctgatcc gtggcggtcg tgttaaagac ctcccgggtg
     4021 ttcgttacca caccgtacgt ggtgcgcttg actgctccgg cgttaaagac cgtaagcagg
     4081 ctcgttccaa gtatggcgtg aagcgtccta aggcttaatg gttctccgtt aagtaaggcc
     4141 aaacgtttta acttaaatgt caaactaaac tcgtagagtt ttggacaatc ctgattattg
     4201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ggtaataact gatataatta
     4321 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     4381 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     4441 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     4501 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     4561 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     4621 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     4681 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     4741 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     4801 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     4861 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     4921 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     4981 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     5041 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     5101 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     5161 cttatatgta gctttcgaca tgatttatct tcgtttcctg caggtttttg ttctgtgcag
     5221 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     5281 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     5341 aaaatttcaa agaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaattg
     5401 aaaagctagc ttatcgatgg gtccttttca tcacgtgcta taaaaataat tataatttaa
     5461 attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa
     5521 atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct
     5581 tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta
     5641 gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata
     5701 tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa
     5761 attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat
     5821 ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa
     5881 attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atctctaaga
     5941 aaccattatt atcatgacat taacctataa aaaaggcctc
//
