LOCUS       V031285                 7065 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031285
VERSION     V031285
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1029..3212
                     /label="Hgf(NM_010427)"
                     /note="Hgf(NM_010427)"
                     /gene="Hgf"
     CDS             3231..3260
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3279..3302
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3322..3338)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3349..3971
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(4120..4708)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    5037..5084
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5316..6110)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(6145..6502)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6504..6608)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6635..7063
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gatgtggggg accaaacttc tgccggtcct gttgctgcag catgtcctcc
     1081 tgcacctcct cctgcttcat gtcgccatcc cctatgcaga aggacagaag aaaagaagaa
     1141 atacacttca tgaatttaaa aagtcagcaa aaactactct taccaaggaa gacccattac
     1201 tgaagattaa aaccaaaaaa gtgaactctg cagatgagtg tgccaacagg tgtatcagga
     1261 acaggggctt tacgttcact tgcaaggcct tcgtttttga taagtcaaga aaacgatgct
     1321 actggtatcc tttcaatagt atgtcaagtg gagtgaaaaa agggtttggc catgaatttg
     1381 acctctatga aaacaaagac tatattagaa actgcatcat tggtaaagga ggcagctata
     1441 aagggacggt atccatcact aagagtggca tcaaatgcca gccttggaat tccatgatcc
     1501 cccatgaaca cagctttttg ccttcgagct atcgcggtaa agacctacag gaaaactact
     1561 gtcgaaatcc tcgaggggaa gaagggggac cctggtgttt cacaagcaat ccagaggtac
     1621 gctacgaagt ctgtgacatt cctcagtgtt cagaagttga atgcatgacc tgcaatggtg
     1681 aaagctacag aggtcccatg gatcacacag aatcaggcaa gacttgtcag cgctgggacc
     1741 agcagacacc acaccggcac aagttcttgc cagaaagata tcccgacaag ggctttgatg
     1801 ataattattg ccgcaatcct gatggcaagc cgaggccatg gtgctacact cttgaccctg
     1861 acaccccttg ggagtattgt gcaattaaaa cgtgcgctca cagtgctgtg aatgagactg
     1921 atgtccctat ggaaacaact gaatgcattc aaggccaagg agaaggttac aggggaacca
     1981 gcaataccat ttggaatgga attccctgtc agcgttggga ttcgcagtac cctcacaagc
     2041 atgatatcac tcccgagaac ttcaaatgca aggaccttag agaaaattat tgccgcaatc
     2101 cagatggggc tgaatcacca tggtgtttta ccactgaccc aaacatccga gttggctact
     2161 gctctcaaat tcccaagtgt gacgtgtcaa gtggacaaga ttgttatcgt ggcaatggga
     2221 aaaattacat gggcaactta tccaaaacaa ggtctggact tacatgttcc atgtgggaca
     2281 agaatatgga ggatttacac cgtcatatct tctgggagcc agatgctagc aaattgaata
     2341 agaattactg ccggaatcct gatgatgatg cccatggacc ttggtgctac acggggaatc
     2401 ctcttattcc ttgggattat tgccctattt cccgttgtga aggagatact acacctacaa
     2461 ttgtcaattt ggaccatcct gtaatatcct gtgccaaaac aaaacaactg cgggttgtaa
     2521 atggcattcc aacacaaaca acagtagggt ggatggttag tttgaaatac agaaataaac
     2581 atatctgtgg aggatcattg ataaaggaaa gttgggttct tactgcaaga caatgttttc
     2641 cagccagaaa caaagacttg aaagactatg aagcttggct tggcatccac gatgttcatg
     2701 agagaggcga ggagaagcgc aagcagatct taaacatttc ccagctggtc tatggtcctg
     2761 aaggctcaga cttggtttta ctgaagcttg ctcgacctgc aatcctggat aactttgtca
     2821 gtacaattga tttacctagt tatggttgta caatccctga aaagaccact tgcagtattt
     2881 acggctgggg ctacactgga ttgatcaacg cggatggttt attacgagta gctcatctgt
     2941 atattatggg gaatgagaaa tgcagtcagc accatcaagg caaggtgact ttgaatgagt
     3001 ctgagttatg tgctggggct gaaaagattg gatcaggacc atgtgaggga gattatggtg
     3061 gcccactcat ttgtgaacaa cacaaaatga gaatggttct tggtgtcatt gttcctggtc
     3121 gtggatgtgc catcccaaat cgtcctggta tttttgttcg agtagcatat tatgcaaaat
     3181 ggatacacaa agtaattttg acatacaagt tgacgcgtac gcggccgctc gagcagaaac
     3241 tcatctcaga agaggatctg gcagcaaatg atatcctgga ttacaaggat gacgacgata
     3301 aggtttaaac ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc
     3361 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3421 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3481 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     3541 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     3601 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     3661 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     3721 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     3781 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa
     3841 ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg
     3901 ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg
     3961 cctgttgaat tgggtacgcg gccagcggcg agcggtatca gctcactcaa aggcggtaat
     4021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4861 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
     4921 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
     4981 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
     5041 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
     5101 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
     5161 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
     5221 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
     5281 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
     5341 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     5401 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     5461 cgatccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     5521 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     5581 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     5641 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     5701 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     5761 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     5821 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     5881 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     5941 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     6001 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     6061 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     6121 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
     6181 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     6241 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     6301 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     6361 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     6421 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     6481 cagctggttc tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag
     6541 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6601 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc
     6661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     6721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     7021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga atttt
//
