LOCUS       V032389                 6255 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032389
VERSION     V032389
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             613..1329
                     /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             1384..2910
                     /label="HARS1(NM_002109)"
                     /note="HARS1(NM_002109)"
                     /gene="HARS1"
     polyA_signal    3043..3164
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3171..3626)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3653..3757
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3759..4116
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4151..4945
                     /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    5177..5224
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5553..6141
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg gggaagcaag
     1381 cttatggcag agcgtgcggc gctggaggag ctggtgaaac ttcagggaga gcgcgtgcga
     1441 ggcctcaagc agcagaaggc cagcgccgag ctgatcgagg aggaggtggc gaaactcctg
     1501 aaactgaagg cacagctggg tcctgatgaa agcaaacaga aatttgtgct caaaaccccc
     1561 aagggcacaa gagactatag tccccggcag atggcagttc gcgagaaggt gtttgacgta
     1621 atcatccgtt gcttcaagcg ccacggtgca gaagtcattg atacacctgt atttgaacta
     1681 aaggaaacac tgatgggaaa gtatggggaa gactccaagc ttatctatga cctgaaggac
     1741 cagggcgggg agctcctgtc ccttcgctat gacctcactg ttccttttgc tcggtatttg
     1801 gcaatgaata aactgaccaa cattaaacgc taccacatag caaaggtata tcggcgggat
     1861 aacccagcca tgacccgtgg ccgataccgg gaattctacc agtgtgattt tgacattgct
     1921 gggaactttg atcccatgat ccctgatgca gagtgcctga agatcatgtg cgagatcctg
     1981 agttcacttc agataggcga cttcctggtc aaggtaaacg atcgacgcat tctagatggg
     2041 atgtttgcta tctgtggtgt ttctgacagc aagttccgta ccatctgctc ctcagtagac
     2101 aagctggaca aggtgtcctg ggaagaggtg aagaatgaga tggtgggaga gaagggcctt
     2161 gcacctgagg tggctgaccg cattggggac tatgtccagc aacatggtgg ggtatccctg
     2221 gtggaacagc tgctccagga tcctaaacta tcccaaaaca agcaggcctt ggagggcctg
     2281 ggagacctga agttgctctt tgagtacctg accctatttg gcattgatga caaaatctcc
     2341 tttgacctga gccttgctcg agggctggat tactacactg gggtgatcta tgaggcagtg
     2401 ctgctacaga ccccagccca ggcaggggaa gagcccctgg gtgtgggcag tgtggctgct
     2461 ggaggacgct atgatgggct agtgggcatg ttcgacccca aagggcgcaa ggtgccatgt
     2521 gtggggctca gcattggggt ggagcggatt ttctccatcg tggaacagag actagaggct
     2581 ttggaggaga agatacggac cacggagaca caggtgcttg tggcatctgc acagaagaag
     2641 ctgctagagg aaagactaaa gcttgtctca gaactgtggg atgctgggat caaggctgag
     2701 ctgctgtaca agaagaaccc aaagctactg aaccagttac agtactgtga ggaggcaggc
     2761 atcccactgg tggctatcat cggcgagcag gaactcaagg atggggtcat caagctccgt
     2821 tcagtgacga gcagggaaga ggtggatgtc cgaagagaag accttgtgga ggaaatcaaa
     2881 aggagaacag gccagcccct ctgcatctgc tgaggatcca ccggatctag ataactgatc
     2941 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3001 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3061 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3121 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     3181 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3241 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3301 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3361 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3421 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3481 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3541 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3601 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3661 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3721 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4021 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4081 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4141 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4201 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4261 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4321 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4381 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4441 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4501 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4561 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4621 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4681 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4741 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4801 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4861 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4921 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4981 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5041 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5101 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5161 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5221 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5281 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5341 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     5701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6241 attaccgcca tgcat
//
