LOCUS       V003531                 4961 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003531
VERSION     V003531
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4961)
  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 4961)
  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 4961)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4961)
  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..4961
                     /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..2533
                     /label="mEos2"
                     /note="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
     misc_feature    2534..2569
                     /label="linker 2"
                     /note="linker 2"
     CDS             2748..3218
                     /gene="dhfrI"
                     /label="Dihydrofolate reductase type 1"
                     /note="Dihydrofolate reductase type 1 from Escherichia
                     coli. Accession#: P00382"
     primer_bind     complement(3449..3465)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(3900..4268)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(4301..4410)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      4570..4958
                     /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 ggttctggcg aattcatgag
     1861 tgcgattaag ccagacatga agatcaaact ccgtatggaa ggcaacgtaa acgggcacca
     1921 ctttgtgatc gacggagatg gtacaggcaa gccttttgag ggaaaacaga gtatggatct
     1981 tgaagtcaaa gagggcggac ctctgccttt tgcctttgat atcctgacca ctgcattcca
     2041 ttacggcaac agggtattcg ccaaatatcc agacaacata caagactatt ttaagcagtc
     2101 gtttcctaag gggtattcgt gggaacgaag cttgactttc gaagacgggg gcatttgcat
     2161 tgccagaaac gacataacaa tggaagggga cactttctat aataaagttc gattttatgg
     2221 taccaacttt cccgccaatg gtccagttat gcagaagaag acgctgaaat gggagccctc
     2281 cactgagaaa atgtatgtgc gtgatggagt gctgacgggt gatattcata tggctttgtt
     2341 gcttgaagga aatgcccatt accgatgtga cttcagaact acttacaaag ctaaggagaa
     2401 gggtgtcaag ttaccaggct accactttgt ggaccactgc attgagattt taagccatga
     2461 caaagattac aacaaggtta agctgtatga gcatgctgtt gctcattctg gattgcctga
     2521 caatgccaga cgaggctctg caggatctgc tgcaggatct ggagaattta gtttaaactc
     2581 tagatgcaag caggatagac ggcatgcacg atttgtaata acagagtgtc ttgtattttt
     2641 agagaaagtc tatttaatac aagtgattat attaattaac ggtaagcatc agcgggtgac
     2701 aaaacgagca tgcttactaa taaaatgtta acctctgagg aagaattgtg aaactatcac
     2761 taatggtagc tatatcgaag aatggagtta tcgggaatgg ccctgatatt ccatggagtg
     2821 ccaaaggtga acagctcctg tttaaagcta ttacctataa ccaatggctg ttggttggac
     2881 gcaagacttt tggatcaatg ggagcattac ccaaccgaaa gtatgcggtc gtaacacgtt
     2941 caagttttac atctgacaat gagaacgtat tgatctttcc atcaattaaa gatgctttaa
     3001 ccaacctaaa gaaaataacg gatcatgtca ttgtttcagg tggtggggag atatacaaaa
     3061 gcctgatcga tcaagtagat acactacata tatctacaat agacatcgag ccggaaggtg
     3121 atgtttactt tcctgaaatc cccagcaatt ttaggccagt ttttacccaa gacttcgcct
     3181 ctaacataaa ttatagttac caaatctggc aaaagggtta acaagtggca gcaacggatt
     3241 cgcaaacctg tcacgccttt tgtgccaaaa gccgcgccag gtttgcgatc cgctgtgcca
     3301 ggcgttaggc gtcatatgaa gatttcggtg atccctgagc aggtggcgga aacattggat
     3361 gctgactgca gcctagttta agactttatt gtcatagttt agatcttttt tgtccagttt
     3421 gagacttttt tgtgtttaaa cagaattcac tggccgtcgt tttacaacgt cgtgactggg
     3481 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3541 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3601 aatggggctc gatcccctcg cgagttggtt cagctgctgc ctgaggctgg acgacctcgc
     3661 ggagttctac cggcagtgca aatccgtcgg catccaggaa accagcagcg gctatccgcg
     3721 catccatgcc cccgaactgc aggagtgggg aggcacgatg gccgctttgg tcggtcttgc
     3781 cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca
     3841 tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat
     3901 ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct
     3961 tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg
     4021 tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc
     4081 agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc
     4141 agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt
     4201 tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca
     4261 tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca
     4321 ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc
     4381 gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag
     4441 tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa
     4501 aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaacggcca cgatgcgtcc
     4561 ggcgtagagg atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat
     4621 gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac
     4681 cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta
     4741 ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa
     4801 ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt ttaaggttta
     4861 acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa
     4921 gcaattttga gtgacacagg aacacttaac ggctgacatg g
//
