LOCUS       V021061                 9715 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021061
VERSION     V021061
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             624..5063
                     /label="CFTR(NM_000492)"
                     /note="CFTR(NM_000492)"
                     /gene="CFTR"
     misc_feature    5074..5660
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5661..6380
                     /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"
     polyA_signal    6503..6624
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(6631..7086)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        7113..7217
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        7219..7576
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7611..8405
                     /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"
     polyA_signal    8637..8684
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      9013..9601
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatgcaga ggtcgcctct ggaaaaggcc agcgttgtct
      661 ccaaactttt tttcagctgg accagaccaa ttttgaggaa aggatacaga cagcgcctgg
      721 aattgtcaga catataccaa atcccttctg ttgattctgc tgacaatcta tctgaaaaat
      781 tggaaagaga atgggataga gagctggctt caaagaaaaa tcctaaactc attaatgccc
      841 ttcggcgatg ttttttctgg agatttatgt tctatggaat ctttttatat ttaggggaag
      901 tcaccaaagc agtacagcct ctcttactgg gaagaatcat agcttcctat gacccggata
      961 acaaggagga acgctctatc gcgatttatc taggcatagg cttatgcctt ctctttattg
     1021 tgaggacact gctcctacac ccagccattt ttggccttca tcacattgga atgcagatga
     1081 gaatagctat gtttagtttg atttataaga agactttaaa gctgtcaagc cgtgttctag
     1141 ataaaataag tattggacaa cttgttagtc tcctttccaa caacctgaac aaatttgatg
     1201 aaggacttgc attggcacat ttcgtgtgga tcgctccttt gcaagtggca ctcctcatgg
     1261 ggctaatctg ggagttgtta caggcgtctg ccttctgtgg acttggtttc ctgatagtcc
     1321 ttgccctttt tcaggctggg ctagggagaa tgatgatgaa gtacagagat cagagagctg
     1381 ggaagatcag tgaaagactt gtgattacct cagaaatgat tgaaaatatc caatctgtta
     1441 aggcatactg ctgggaagaa gcaatggaaa aaatgattga aaacttaaga caaacagaac
     1501 tgaaactgac tcggaaggca gcctatgtga gatacttcaa tagctcagcc ttcttcttct
     1561 cagggttctt tgtggtgttt ttatctgtgc ttccctatgc actaatcaaa ggaatcatcc
     1621 tccggaaaat attcaccacc atctcattct gcattgttct gcgcatggcg gtcactcggc
     1681 aatttccctg ggctgtacaa acatggtatg actctcttgg agcaataaac aaaatacagg
     1741 atttcttaca aaagcaagaa tataagacat tggaatataa cttaacgact acagaagtag
     1801 tgatggagaa tgtaacagcc ttctgggagg agggatttgg ggaattattt gagaaagcaa
     1861 aacaaaacaa taacaataga aaaacttcta atggtgatga cagcctcttc ttcagtaatt
     1921 tctcacttct tggtactcct gtcctgaaag atattaattt caagatagaa agaggacagt
     1981 tgttggcggt tgctggatcc actggagcag gcaagacttc acttctaatg gtgattatgg
     2041 gagaactgga gccttcagag ggtaaaatta agcacagtgg aagaatttca ttctgttctc
     2101 agttttcctg gattatgcct ggcaccatta aagaaaatat catctttggt gtttcctatg
     2161 atgaatatag atacagaagc gtcatcaaag catgccaact agaagaggac atctccaagt
     2221 ttgcagagaa agacaatata gttcttggag aaggtggaat cacactgagt ggaggtcaac
     2281 gagcaagaat ttctttagca agagcagtat acaaagatgc tgatttgtat ttattagact
     2341 ctccttttgg atacctagat gttttaacag aaaaagaaat atttgaaagc tgtgtctgta
     2401 aactgatggc taacaaaact aggattttgg tcacttctaa aatggaacat ttaaagaaag
     2461 ctgacaaaat attaattttg catgaaggta gcagctattt ttatgggaca ttttcagaac
     2521 tccaaaatct acagccagac tttagctcaa aactcatggg atgtgattct ttcgaccaat
     2581 ttagtgcaga aagaagaaat tcaatcctaa ctgagacctt acaccgtttc tcattagaag
     2641 gagatgctcc tgtctcctgg acagaaacaa aaaaacaatc ttttaaacag actggagagt
     2701 ttggggaaaa aaggaagaat tctattctca atccaatcaa ctctatacga aaattttcca
     2761 ttgtgcaaaa gactccctta caaatgaatg gcatcgaaga ggattctgat gagcctttag
     2821 agagaaggct gtccttagta ccagattctg agcagggaga ggcgatactg cctcgcatca
     2881 gcgtgatcag cactggcccc acgcttcagg cacgaaggag gcagtctgtc ctgaacctga
     2941 tgacacactc agttaaccaa ggtcagaaca ttcaccgaaa gacaacagca tccacacgaa
     3001 aagtgtcact ggcccctcag gcaaacttga ctgaactgga tatatattca agaaggttat
     3061 ctcaagaaac tggcttggaa ataagtgaag aaattaacga agaagactta aaggagtgct
     3121 tttttgatga tatggagagc ataccagcag tgactacatg gaacacatac cttcgatata
     3181 ttactgtcca caagagctta atttttgtgc taatttggtg cttagtaatt tttctggcag
     3241 aggtggctgc ttctttggtt gtgctgtggc tccttggaaa cactcctctt caagacaaag
     3301 ggaatagtac tcatagtaga aataacagct atgcagtgat tatcaccagc accagttcgt
     3361 attatgtgtt ttacatttac gtgggagtag ccgacacttt gcttgctatg ggattcttca
     3421 gaggtctacc actggtgcat actctaatca cagtgtcgaa aattttacac cacaaaatgt
     3481 tacattctgt tcttcaagca cctatgtcaa ccctcaacac gttgaaagca ggtgggattc
     3541 ttaatagatt ctccaaagat atagcaattt tggatgacct tctgcctctt accatatttg
     3601 acttcatcca gttgttatta attgtgattg gagctatagc agttgtcgca gttttacaac
     3661 cctacatctt tgttgcaaca gtgccagtga tagtggcttt tattatgttg agagcatatt
     3721 tcctccaaac ctcacagcaa ctcaaacaac tggaatctga aggcaggagt ccaattttca
     3781 ctcatcttgt tacaagctta aaaggactat ggacacttcg tgccttcgga cggcagcctt
     3841 actttgaaac tctgttccac aaagctctga atttacatac tgccaactgg ttcttgtacc
     3901 tgtcaacact gcgctggttc caaatgagaa tagaaatgat ttttgtcatc ttcttcattg
     3961 ctgttacctt catttccatt ttaacaacag gagaaggaga aggaagagtt ggtattatcc
     4021 tgactttagc catgaatatc atgagtacat tgcagtgggc tgtaaactcc agcatagatg
     4081 tggatagctt gatgcgatct gtgagccgag tctttaagtt cattgacatg ccaacagaag
     4141 gtaaacctac caagtcaacc aaaccataca agaatggcca actctcgaaa gttatgatta
     4201 ttgagaattc acacgtgaag aaagatgaca tctggccctc agggggccaa atgactgtca
     4261 aagatctcac agcaaaatac acagaaggtg gaaatgccat attagagaac atttccttct
     4321 caataagtcc tggccagagg gtgggcctct tgggaagaac tggatcaggg aagagtactt
     4381 tgttatcagc ttttttgaga ctactgaaca ctgaaggaga aatccagatc gatggtgtgt
     4441 cttgggattc aataactttg caacagtgga ggaaagcctt tggagtgata ccacagaaag
     4501 tatttatttt ttctggaaca tttagaaaaa acttggatcc ctatgaacag tggagtgatc
     4561 aagaaatatg gaaagttgca gatgaggttg ggctcagatc tgtgatagaa cagtttcctg
     4621 ggaagcttga ctttgtcctt gtggatgggg gctgtgtcct aagccatggc cacaagcagt
     4681 tgatgtgctt ggctagatct gttctcagta aggcgaagat cttgctgctt gatgaaccca
     4741 gtgctcattt ggatccagta acataccaaa taattagaag aactctaaaa caagcatttg
     4801 ctgattgcac agtaattctc tgtgaacaca ggatagaagc aatgctggaa tgccaacaat
     4861 ttttggtcat agaagagaac aaagtgcggc agtacgattc catccagaaa ctgctgaacg
     4921 agaggagcct cttccggcaa gccatcagcc cctccgacag ggtgaagctc tttccccacc
     4981 ggaactcaag caagtgcaag tctaagcccc agattgctgc tctgaaagag gagacagaag
     5041 aagaggtgca agatacaagg ctttagggat ccgcccctct ccctcccccc cccctaacgt
     5101 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     5161 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     5221 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     5281 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     5341 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     5401 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     5461 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     5521 ccattgtatg ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag
     5581 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     5641 atgataatat ggccacaacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     5701 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     5761 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     5821 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     5881 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     5941 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     6001 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     6061 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     6121 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     6181 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     6241 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     6301 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     6361 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     6421 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6481 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     6541 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6601 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     6661 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     6721 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6781 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6841 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6901 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6961 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     7021 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     7081 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7201 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     7261 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     7321 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     7381 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     7441 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     7501 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7561 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     7621 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     7681 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     7741 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     7801 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     7861 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     7921 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     7981 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     8041 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     8101 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     8161 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     8221 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     8281 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     8341 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     8401 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     8461 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     8521 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     8581 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     8641 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     8701 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     8761 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     8821 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     8881 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8941 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     9001 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     9061 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     9121 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     9181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     9241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     9301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     9361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     9421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     9481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     9541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     9601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     9661 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
