LOCUS       V003533                 4391 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003533
VERSION     V003533
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4391)
  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 4391)
  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 4391)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4391)
  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..4391
                     /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             1856..1885
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             1895..1924
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label="Myc"
                     /translation="EQKLISEEDL"
     CDS             1934..1963
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label="Myc"
                     /translation="EQKLISEEDL"
     misc_feature    1970..1999
                     /label="linker 2"
                     /note="linker 2"
     CDS             2178..2648
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     primer_bind     complement(2879..2895)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3330..3698)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(3731..3840)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      4000..4388
                     /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 ggttctggct acatggaaca
     1861 aaagttgatt tctgaggagg acttgaatga aatggagcaa aaactcattt ctgaggaaga
     1921 cttgaatgaa atggagcaaa agctcatttc tgaagaagac ctcaatgaag gctctgcagg
     1981 atctgctgca ggaggagcta gtttaaactc tagatgcaag caggatagac ggcatgcacg
     2041 atttgtaata acagagtgtc ttgtattttt agagaaagtc tatttaatac aagtgattat
     2101 attaattaac ggtaagcatc agcgggtgac aaaacgagca tgcttactaa taaaatgtta
     2161 acctctgagg aagaattgtg aaactatcac taatggtagc tatatcgaag aatggagtta
     2221 tcgggaatgg ccctgatatt ccatggagtg ccaaaggtga acagctcctg tttaaagcta
     2281 ttacctataa ccaatggctg ttggttggac gcaagacttt tggatcaatg ggagcattac
     2341 ccaaccgaaa gtatgcggtc gtaacacgtt caagttttac atctgacaat gagaacgtat
     2401 tgatctttcc atcaattaaa gatgctttaa ccaacctaaa gaaaataacg gatcatgtca
     2461 ttgtttcagg tggtggggag atatacaaaa gcctgatcga tcaagtagat acactacata
     2521 tatctacaat agacatcgag ccggaaggtg atgtttactt tcctgaaatc cccagcaatt
     2581 ttaggccagt ttttacccaa gacttcgcct ctaacataaa ttatagttac caaatctggc
     2641 aaaagggtta acaagtggca gcaacggatt cgcaaacctg tcacgccttt tgtgccaaaa
     2701 gccgcgccag gtttgcgatc cgctgtgcca ggcgttaggc gtcatatgaa gatttcggtg
     2761 atccctgagc aggtggcgga aacattggat gctgactgca gcctagttta agactttatt
     2821 gtcatagttt agatcttttt tgtccagttt gagacttttt tgtgtttaaa cagaattcac
     2881 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2941 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3001 cttcccaaca gttgcgcagc ctgaatggcg aatggggctc gatcccctcg cgagttggtt
     3061 cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca aatccgtcgg
     3121 catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc aggagtgggg
     3181 aggcacgatg gccgctttgg tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct
     3241 ccgcgaagtc gctcttcttg atggagcgca tggggacgtg cttggcaatc acgcgcaccc
     3301 cccggccgtt ttagcggcta aaaaagtcat ggctctgccc tcgggcggac cacgcccatc
     3361 atgaccttgc caagctcgtc ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg
     3421 gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc
     3481 aggcggccca ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg
     3541 cccgtgattt tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc
     3601 gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt
     3661 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     3721 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     3781 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     3841 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     3901 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     3961 tatgcagcgg aaaacggcca cgatgcgtcc ggcgtagagg atctgaagat cagcagttca
     4021 acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg
     4081 tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgg
     4141 ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta
     4201 atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat
     4261 atataaggct tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa
     4321 cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac
     4381 ggctgacatg g
//
