LOCUS       V020283                 7084 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020283
VERSION     V020283
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..2996
                     /label="CLCN3(NM_001243374)"
                     /note="CLCN3(NM_001243374)"
                     /gene="CLCN3"
     CDS             3030..3749
                     /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    3872..3993
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4000..4455)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4482..4586
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4588..4945
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4980..5774
                     /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    6006..6053
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6382..6970
                     /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 accatggatg cttcttccga cccttatttg ccttatgacg
      661 ggggaggaga cagtattccc ctgagggaat tacataaaag aggaactcat tatacaatga
      721 caaatggagg cagcattaac agttctacac atttactgga tcttttggat gaaccaattc
      781 caggtgttgg tacatatgat gatttccata ctattgattg ggtgcgagaa aaatgtaaag
      841 acagagaaag gcatagacgg atcaacagca aaaagaaaga atcagcatgg gaaatgacaa
      901 aaagtttgta tgatgcgtgg tcaggatggc tagtagtaac actaacagga ttggcatcag
      961 gggcactggc cggattaata gacattgctg ccgattggat gactgaccta aaggagggca
     1021 tttgccttag tgcgttgtgg tacaaccacg aacagtgctg ttggggatct aatgaaacaa
     1081 catttgaaga gagggataaa tgtccacagt ggaaaacatg ggcagaatta atcataggtc
     1141 aagcagaggg tcctggttct tatatcatga actacataat gtacatcttc tgggccttga
     1201 gttttgcctt tcttgcagtt tccctggtaa aggtatttgc tccatatgcc tgtggctctg
     1261 gaattccaga gattaaaact attttaagtg gattcatcat cagaggttac ttgggaaaat
     1321 ggactttaat gattaaaacc atcacattag tcctggctgt ggcatcaggt ttgagtttag
     1381 gaaaagaagg tcccctggta catgttgcct gttgctgcgg aaatatcttt tcctacctct
     1441 ttccaaagta tagcacaaac gaagctaaaa aaagggaggt gctatcagct gcctcagctg
     1501 caggggtttc tgtagctttt ggtgcaccaa ttggaggagt tctttttagc ctggaagagg
     1561 ttagctatta ttttcctctc aaaactttat ggagatcatt ttttgctgct ttagtggctg
     1621 catttgtttt gaggtccatc aatccatttg gtaacagccg tctggtcctt ttttatgtgg
     1681 agtatcatac accatggtac ctttttgaac tgtttccttt tattcttcta ggggtatttg
     1741 gagggctttg gggagccttt ttcattaggg caaatattgc ctggtgtcgt cgacgcaagt
     1801 ccacgaaatt tggaaagtat cccgttctgg aagtcattat tgttgcagcc attactgctg
     1861 tgatagcctt ccctaatcca tacactaggc taaacaccag tgaactgatc aaagagcttt
     1921 ttacagactg tggtcccctg gaatcctctt ctctttgtga ctacagaaat gacatgaatg
     1981 ccagtaaaat tgtcgatgac attcctgatc gtccagcagg cattggagta tattcagcta
     2041 tatggcagtt atgcctggca ctcatattta aaatcataat gacagtattc acttttggca
     2101 tcaaggttcc atcaggcttg ttcatcccca gcatggccat tggagcgatc gcaggaagga
     2161 ttgtggggat tgcggtggag cagcttgcct actatcacca cgactggttt atctttaagg
     2221 agtggtgtga ggtcggggct gattgcatta cacctggcct ttatgccatg gttggtgctg
     2281 ctgcatgctt aggtggtgtg acaagaatga ctgtctccct ggtggttatt gtttttgagc
     2341 ttactggagg cttggaatat attgttcccc ttatggctgc agtcatgacc agtaaatggg
     2401 ttggagatgc ctttggcagg gaaggcattt atgaagcaca catccgatta aatggatacc
     2461 ctttcttgga tgcaaaagaa gaattcactc ataccaccct ggctgctgac gttatgagac
     2521 ctcgaaggaa tgatcctccc ttagctgtcc tgacacagga caatatgaca gtggatgata
     2581 tagaaaacat gattaatgaa accagctaca atggatttcc tgtcataatg tcaaaagaat
     2641 ctcagagatt agtgggattt gccctcagaa gagacctgac aattgcaata gaaagtgcca
     2701 ggaaaaaaca agaaggtatc gttggcagtt ctcgggtgtg ttttgcacag cacaccccat
     2761 ctcttccagc agaaagtcct cggccattga agcttcgaag cattcttgac atgagccctt
     2821 ttacagtgac agaccacacc ccaatggaga tcgtggtgga tattttccga aagctgggac
     2881 tgaggcagtg ccttgtaact cacaatgggc gcctccttgg cattataaca aaaaaagata
     2941 tcctccggca tatggcccag acggcaaacc aagaccccgc ttcaataatg ttcaacggag
     3001 gagggggaag cgatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     3061 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3121 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3181 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3241 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3301 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3361 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3421 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3481 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3541 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3601 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3661 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3721 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca
     3781 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3841 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3901 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3961 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4021 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4081 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4141 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4201 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4261 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4321 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4381 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4441 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4501 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4561 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4621 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4681 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4741 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4801 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4861 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4921 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4981 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5041 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5101 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5161 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5221 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5281 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5341 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5401 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5461 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5521 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5581 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5641 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5701 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5761 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5821 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5881 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5941 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6001 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6061 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6121 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6181 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6241 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6301 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6361 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6421 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6481 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6541 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6601 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6661 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6721 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6781 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6841 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6901 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6961 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7021 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7081 gcat
//
