LOCUS       V003532                 4985 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003532
VERSION     V003532
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4985)
  AUTHORS   Zordan RE, Beliveau BJ, Trow JA, Craig NL, Cormack BP.
  TITLE     Avoiding the ends: internal epitope tagging of proteins using
            transposon Tn7
  JOURNAL   Genetics 200 (1), 47-58 (2015)
   PUBMED   25745023
REFERENCE   2  (bases 1 to 4985)
  AUTHORS   Zordan RE, Cormack BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2015) Molecular Biology and Genetics, Johns
            Hopkins School of Medicine, 725 N Wolfe St., Hunterian 609,
            Baltimore, MD 21205, USA
REFERENCE   3  (bases 1 to 4985)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4985)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics";
            date: "2015"; volume: "200"; issue: "1"; pages: "47-58"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-JAN-2015) Molecular Biology and Genetics, Johns Hopkins School
            of Medicine, 725 N Wolfe St., Hunterian 609, Baltimore, MD 21205,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             361..1152
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     protein_bind    1430..1451
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1466..1496
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1504..1520
                     /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)."
     primer_bind     1528..1544
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    1820..1849
                     /label="linker 1"
                     /note="linker 1"
     CDS             1850..2563
                     /label="yeGFP"
                     /note="yeast-enhanced green fluorescent protein"
     misc_feature    2564..2593
                     /label="linker 2"
                     /note="linker 2"
     CDS             2772..3242
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     primer_bind     complement(3473..3489)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3924..4292)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4325..4434)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      4594..4982
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gaattagctt cacgctgccg caagcactca gggcgcaagg gctgctaaag gaagcggaac
       61 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
      121 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
      181 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
      241 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
      301 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
      361 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
      421 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      481 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      541 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      601 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
      661 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
      721 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
      781 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
      841 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
      901 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
      961 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     1021 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     1081 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     1141 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     1201 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     1261 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     1321 cccacccccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     1381 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     1441 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1501 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     1561 ttgcatgcct gcaggtcgac tctagctaga ggatcctgtg gccggccaat aaagtcttaa
     1621 actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag aatagttgta
     1681 aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag
     1741 caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggggtcg
     1801 acggcgcgcc ggccggcctg gatccgctgg ctccgctgct ggttctggca tgtctaaagg
     1861 tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg
     1921 tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt
     1981 aaaatttatt tgtactactg gtaaattgcc ggttccatgg ccaaccttag tcactacttt
     2041 cggttatggt gttcaatgtt ttgctagata cccagatcat atgaaacaac atgacttttt
     2101 caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg
     2161 taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga
     2221 attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa
     2281 ctataactct cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa
     2341 cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca
     2401 aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact tatccactca
     2461 atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt
     2521 tactgctgct ggtattaccc atggtatgga tgaattgtac aaaggctctg caggatctgc
     2581 tgcaggagga gctagtttaa actctagatg caagcaggat agacggcatg cacgatttgt
     2641 aataacagag tgtcttgtat ttttagagaa agtctattta atacaagtga ttatattaat
     2701 taacggtaag catcagcggg tgacaaaacg agcatgctta ctaataaaat gttaacctct
     2761 gaggaagaat tgtgaaacta tcactaatgg tagctatatc gaagaatgga gttatcggga
     2821 atggccctga tattccatgg agtgccaaag gtgaacagct cctgtttaaa gctattacct
     2881 ataaccaatg gctgttggtt ggacgcaaga cttttggatc aatgggagca ttacccaacc
     2941 gaaagtatgc ggtcgtaaca cgttcaagtt ttacatctga caatgagaac gtattgatct
     3001 ttccatcaat taaagatgct ttaaccaacc taaagaaaat aacggatcat gtcattgttt
     3061 caggtggtgg ggagatatac aaaagcctga tcgatcaagt agatacacta catatatcta
     3121 caatagacat cgagccggaa ggtgatgttt actttcctga aatccccagc aattttaggc
     3181 cagtttttac ccaagacttc gcctctaaca taaattatag ttaccaaatc tggcaaaagg
     3241 gttaacaagt ggcagcaacg gattcgcaaa cctgtcacgc cttttgtgcc aaaagccgcg
     3301 ccaggtttgc gatccgctgt gccaggcgtt aggcgtcata tgaagatttc ggtgatccct
     3361 gagcaggtgg cggaaacatt ggatgctgac tgcagcctag tttaagactt tattgtcata
     3421 gtttagatct tttttgtcca gtttgagact tttttgtgtt taaacagaat tcactggccg
     3481 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3541 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3601 aacagttgcg cagcctgaat ggcgaatggg gctcgatccc ctcgcgagtt ggttcagctg
     3661 ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg tcggcatcca
     3721 ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt ggggaggcac
     3781 gatggccgct ttggtcggtc ttgccttgct cgtcggtgat gtacttcacc agctccgcga
     3841 agtcgctctt cttgatggag cgcatgggga cgtgcttggc aatcacgcgc accccccggc
     3901 cgttttagcg gctaaaaaag tcatggctct gccctcgggc ggaccacgcc catcatgacc
     3961 ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca gggcgaggat cgtggcatca
     4021 ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg
     4081 cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac gacgcccgtg
     4141 attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc ggccgccgcc
     4201 gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat aggtgggctg
     4261 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     4321 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     4381 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     4441 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     4501 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     4561 gcggaaaacg gccacgatgc gtccggcgta gaggatctga agatcagcag ttcaacctgt
     4621 tgatagtacg tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa
     4681 gctctcatgt ttaacgaact aaaccctcat ggctaacgta ctaagctctc atggctaacg
     4741 tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa
     4801 atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa
     4861 ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat gtaacgcact
     4921 gagaagccct tagagcctct caaagcaatt ttgagtgaca caggaacact taacggctga
     4981 catgg
//
