LOCUS       V037815                10858 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037815
VERSION     V037815
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             991..1707
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1771..7005
                     /label="Tjp1(NM_009386)"
                     /note="Tjp1(NM_009386)"
                     /gene="Tjp1"
     CDS             7024..7053
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             7072..7095
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(7115..7131)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    7142..7764
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(7913..8501)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    8830..8877
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(9109..9903)
                     /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(9938..10295)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(10297..10401)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      10428..10856
                     /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 attcgccacc atggtgagca agggcgagga gctgttcacc
     1021 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1081 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1141 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1201 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1261 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1321 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1381 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1441 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1501 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1561 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1621 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1681 actctcggca tggacgagct gtacaagggc ggtggaggca gtggtggagg cggtagcaag
     1741 gtaccgagga gatctgccgc cgcgatcgcc atgtccgcca gggccgcggc cgctaagagc
     1801 acagcaatgg aggaaacagc tatatgggaa cagcacacag tgacgcttca cagggctcct
     1861 gggtttggat ttggaattgc aatctctggt ggaagagata atcctcattt tcagagtggg
     1921 gaaacctcca tagtgatttc tgatgtgtta aaaggagggc cagctgaagg acagctacag
     1981 gaaaatgacc gagttgcaat ggttaacgga gtttcaatgg ataacgttga acatgctttt
     2041 gctgttcagc agctaaggaa gagtgggaaa aacgcaaaaa ttactatccg aaggaagaag
     2101 aaagttcaga tccctgtaag tcacccagat cctgagccgg tgtctgataa tgaagacgat
     2161 agttatgacg aagaagtgca tgacccaaga gctggccgcg gtgctttagc gaacagaagg
     2221 agcgagaaga gctgggcaag ggataggagt gcaagcaggg agaggagcct gtcccctcgc
     2281 tcggacaggc ggtccgtggc ctccagtcag cccgcaaagc ccaccaaggt cacactggtg
     2341 aagtctcgga aaaatgaaga atatggtctt cgattggcca gccatatatt tgtaaaggaa
     2401 atttcacaag atagtttggc agcaagagat ggtaacattc aagaagggga tgttgtcttg
     2461 aagataaatg gtactgtgac agaaaatatg tcattgacag atgcaaaaac actgatagaa
     2521 aggtctaaag gcaagttaaa aatggtagtg caaagagatg agcgggctac cttactgaac
     2581 gtccctgacc tttcggatag tatccattct gctaatgcct cggaaagaga tgacatttca
     2641 gaaattcagt cactagcatc agaccattca ggtcgctcgc atgacaggcc accccgccgc
     2701 agccagtcac gatctcctga ccaacgttca gagccctccg atcattccac gcagtctcca
     2761 cagcagccca gcaatggcag tctccggagc agagaggaag agcgaatgtc taaacctggg
     2821 gccatctcaa ctcctgtaaa acatgtagac gatcatccac ccaaagcagt ggaagaagtt
     2881 acagttgaga aaaatgagaa gcagacgccc actcttccag aaccgaaacc tgtgtatgct
     2941 caagttggac aaccagatgt ggatttaccc gtcagccctt ctgatggtgc tctgcctaat
     3001 tcagctcatg aagacgggat acttaggccc agcatgaaac tggtaaaatt cagaaaagga
     3061 gatagtgtgg gtttgcgact agctggtgga aatgatgtcg gaatatttgt agctggcgtt
     3121 ctagaagata gccctgcagc caaagaaggc ttagaggaag gtgatcaaat tctcagggtg
     3181 aacaatgtag atttcacaaa tatcataagg gaagaggccg tccttttcct ccttgacctc
     3241 cctaaaggtg aagaagtgac catactggct cagaagaaga aggacgttta tcgccgcatt
     3301 gtagaatcag atgtaggaga ttcattctat attagaacgc attttgaata tgaaaaagaa
     3361 tctccttacg gacttagttt taacaaagga gaggtgttcc gggtcgtgga tactttatac
     3421 aatggaaagc tgggctcttg gcttgccatt cgaattggca aaaatcataa ggaggtagaa
     3481 cgaggcatca tccctaataa gaacagagct gaacagttag ccagtgtaca gtacacactt
     3541 ccaaagacag cgggtggtga tcgggcagac ttctggaggt ttcgaggtct tcgcagctcc
     3601 aagagaaatc ttcgaaaaag cagagaggac ttgtcagctc agccagttca aactaagttc
     3661 ccagcttatg aaagggttgt tcttcgagaa gctggattcc taagacctgt aaccatcttt
     3721 ggaccaatag ctgatgttgc cagagaaaag ttggcaagag aggagccaga tatctatcag
     3781 attgcaaaaa gtgaaccacg agatgctggg actgaccatc gtagctctgg catcattcgc
     3841 cttcatacaa taaagcaaat catagatcag gataaacatg ctttattaga tgtaacgcca
     3901 aatgcggttg atcgtcttaa ttatgcgcag tggtatccaa ttgttgtgtt ccttaaccct
     3961 gactctaagc aaggtgtaaa aacaatgagg atgaggttgt gtccggagtc tcggaaaagc
     4021 gccaggaagc tatatgaacg ctctcataag cttcgtaaga acaatcacca tctcttcaca
     4081 actacaatta acttaaactc aatgaatgat ggttggtacg gtgccctgaa agaagcgatt
     4141 cagcagcaac agaaccagct ggtgtgggtc tctgagggga aggcggatgg tgctacaagt
     4201 gatgaccttg atttgcatga cgatcgtctg tcctacctgt cagccccagg tagtgagtac
     4261 tcaatgtata gcacggacag tagacacact tctgactatg aagacacaga tacagaaggc
     4321 ggggcctaca ctgatcaaga actagatgaa actcttaatg atgaggtggg gactcccccg
     4381 gagtctgcca ttacacggtc ctctgagcct gtaagagagg attcctctgg aatgcatcat
     4441 gaaaaccaga cataccctcc ttactcacca caagcgcagc cacaagctat tcatagaata
     4501 gactcccctg gacttaagcc agcctctcaa cagaaagcag aagcctcatc tccagtccct
     4561 tacctttcgc ctgaaacaac cccagcatca tcagcctctg cagttaatca taatgtcagt
     4621 gtaactaatg tcagcctgga ggagcctgcc ccagcccctc ccacctcgca cgcatcacag
     4681 cctggttgtt taggagcacc aagtgctgag gcagctcacg tagtgctcag aggtgaagga
     4741 ccaccattgc cgccgcatgc agacccagca aaggtgtaca ggaaggagcc gtattctgaa
     4801 gaaatgatga gacaaaacca tattttaaaa caaccagctc ttggtcaccc agggcagagg
     4861 ccagataaag agccaaatct agcctatgaa ccccaacttc catatataga aaaacaagcc
     4921 agcagagacc ttgagcagcc gtcatacagg tatgaggtct caagctacac agaccagttt
     4981 tctcggaact atgaccatcg cctacggttt gaagatcgaa tccctaccta tgaagaccag
     5041 tggtcatatt atgatgacaa acagccctac caacctcggc cttttgagaa tcagcatccc
     5101 cgagacctgg actccagaca acatcccgaa gaggcttcag aacgaggtta tttccagcgt
     5161 tttgaagagc cagcccctct gtcgtacgac agtagaacac gctatgagca gctgcctcga
     5221 acctctactc tacgacatga agagcagcca gcccctgcat atgaggtgca caacaggtac
     5281 aggccagagg cacagcccta ttcttcaaca ggccctaagt catctgagcc caagcagtac
     5341 tttgaccagt acccgcgaag ttatgagcaa gttccaccac caggctttac ctccaaaaca
     5401 ggccattacg agcctctcca tggtgctgca gttgtccctc ctctgatacc ttcctctcaa
     5461 caaaagccag aagtcctgcc ctcggctacc aaaccacagc ctccaccccc aaccctaact
     5521 gaggaggagg aggatccagc aatgaaacca cagtctgtgc tcaccagagt caaaatgttt
     5581 gaaaacaaaa gatctgcgtc tttggagaac aagaaagatg tgaatgacac tgccagcttt
     5641 aagcctccgg aagtagcatc taaacctcca ggtgcttctc ttgctggccc taaacctgtc
     5701 cctcagagtc agtttagtga gcacgacaaa acgctctaca ggctcccaga gcctcagaaa
     5761 cctcaagtga agccacccga agatattgtt cgatcaaatc attacgaccc tgaagaggat
     5821 gaagaatatt accggaaaca gctctcctac tttgaccgaa gaagttttga gagcaagcct
     5881 tctgcacatc ttcctgctgg ccatcactca gagcctgcta agccagtcca ttctcagagc
     5941 cagcccaact tctctagtta ctcttcaaag ggaaaacccg aaactgatgc tgtggataga
     6001 tcattcagtg agaaacgtta tgatccagcc caggccacgc ctcctcctcc tccgttgccc
     6061 tcacagtaca gccagccagc tccacctctg tccagctctt ctctccacat acattccaag
     6121 ggcgcccagg gtgaaggcaa ctcagtatca ttggattttc agaactcata tatgtccaaa
     6181 ccagacccac ccccatctca gagtaaacca gcaactttca gaccaccaac tcgagaagac
     6241 ccccctcaga ctttctatcc gcagaaaagt ttcccagaca aagctccagt taacggagct
     6301 gagcagactc agaaaaccat cactccggtg tacaatcgat tcacaccaaa gccgtacaca
     6361 agttctgccc ggccatttga acgcaaattt gaaagtccga agttcaacca taatcttctg
     6421 ccaagtgaaa ctgtacataa acctgaattg tcttcaaaaa ctcccacttc cccaaaaact
     6481 cttatgaaag ctcatagttc aacacagcct ccagagtttg acagtggagt tgaaactttc
     6541 tctgttcaca cagataagcc taaatatcaa atgaataata tcagcaccat gcctaaagct
     6601 gtccctgtga gtccttcagc tgtggaagaa gatgaagatg aggatggtca tactgtagtg
     6661 gctacagctc gtggcatttt taacagcaat ggtggtgtgt tgagttccat agaaaccggt
     6721 gttagtataa ttatcccaca aggagccatt cctgaaggaa ttgagcaaga aatctatttc
     6781 aaagtctgca gagacaatag cattctccca cctttagata aagagaaagg tgaaactctg
     6841 ctgagccccc tagtgatgtg tgggccccat ggcctcaagt tcctgaagcc cgtggagcta
     6901 cgcttgccac actgtgaccc taaaacctgg caaaacaagt gtcttcctgg agatccgaat
     6961 taccttgttg gagccaactg tgtttctgtc ctgattgacc actttacgcg tacgcggccg
     7021 ctcgagcaga aactcatctc agaagaggat ctggcagcaa atgatatcct ggattacaag
     7081 gatgacgacg ataaggttta aacggccggc cgcggtcata gctgtttcct gaacagatcc
     7141 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     7201 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     7261 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     7321 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     7381 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     7441 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     7501 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     7561 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     7621 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca
     7681 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc
     7741 cactcagtag atgcctgttg aattgggtac gcggccagcg gcgagcggta tcagctcact
     7801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8641 aaagtatata tgagtaacct gaggctatgg cagggcctgc cgccccgacg ttggctgcga
     8701 gccctgggcc ttcacccgaa cttggggggt ggggtgggga aaaggaagaa acgcgggcgt
     8761 attggcccca atggggtctc ggtggggtat cgacagagtg ccagccctgg gaccgaaccc
     8821 cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca
     8881 tagcgcgggt tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt
     8941 gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga
     9001 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
     9061 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
     9121 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     9181 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     9241 tatgtcctga tagcgatccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     9301 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     9361 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     9421 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
     9481 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     9541 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     9601 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     9661 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     9721 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     9781 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     9841 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     9901 catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa
     9961 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
    10021 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
    10081 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
    10141 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
    10201 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
    10261 gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc aatattattg
    10321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
    10441 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
    10501 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
    10561 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
    10621 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
    10681 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
    10741 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
    10801 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaatttt
//
