LOCUS       V026976                 6779 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026976
VERSION     V026976
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..2096
                     /label="CYP2F1(NM_000774)"
                     /note="CYP2F1(NM_000774)"
                     /gene="CYP2F1"
     misc_feature    2138..2724
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             2725..3444
                     /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    3567..3688
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3695..4150)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4177..4281
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4283..4640
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4675..5469
                     /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    5701..5748
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6077..6665
                     /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 accatggaca gcataagcac agccatctta ctcctgctcc
      661 tggctctcgt ctgtctgctc ctgaccctaa gctcaagaga taagggaaag ctgcctccgg
      721 gacccagacc cctctcaatc ctgggaaacc tgctgctgct ttgctcccaa gacatgctga
      781 cttctctcac taagctgagc aaggagtatg gctccatgta cacagtgcac ctgggaccca
      841 ggcgggtggt ggtcctcagc gggtaccaag ctgtgaagga ggccctggtg gaccagggag
      901 aggagtttag tggccgcggt gactaccctg cctttttcaa ctttaccaag ggcaatggca
      961 tcgccttctc cagtggggat cgatggaagg tcctgagaca gttctctatc cagattctac
     1021 ggaatttcgg gatggggaag agaagcattg aggagcgaat cctagaggag ggcagcttcc
     1081 tgctggcgga gctgcggaaa actgaaggcg agccctttga ccccacgttt gtgctgagtc
     1141 gctcagtgtc caacattatc tgttccgtgc tcttcggcag ccgcttcgac tatgatgatg
     1201 agcgtctgct caccattatc cgccttatca atgacaactt ccaaatcatg agcagcccct
     1261 ggggcgagtt gtacgacatc ttcccgagcc tcctggactg ggtgcctggg ccgcaccaac
     1321 gcatcttcca gaacttcaag tgcctgagag acctcatcgc ccacagcgtc cacgaccacc
     1381 aggcctcgct agaccccaga tctccccggg acttcatcca gtgcttcctc accaagatgg
     1441 cagaggagaa ggaggaccca ctgagccact tccacatgga taccctgctg atgaccacac
     1501 ataacctgct ctttggcggc accaagacgg tgagcaccac gctgcaccac gccttcctgg
     1561 cactcatgaa gtacccaaaa gttcaagccc gcgtgcagga ggagatcgac ctcgtggtgg
     1621 gacgcgcgcg gctgccggcg ctgaaggacc gcgcggccat gccttacaca gacgcggtga
     1681 tccacgaggt gcagcgcttt gcagacatca tccccatgaa cttgccgcac cgcgtcacta
     1741 gggacacggc ctttcgcggc ttcctgatac ccaagggcac cgatgtcatc accctcctta
     1801 acaccgtcca ctacgacccc agccagttcc tgacgcccca ggagttcaac cccgagcatt
     1861 ttttggatgc caatcagtcc ttcaagaaga gtccagcctt catgcccttc tcagctgggc
     1921 gccgtctgtg cctgggagag tcgctggcgc gcatggagct ctttctgtac ctcaccgcca
     1981 tcctgcagag cttttcgctg cagccgctgg gtgcgcccga ggacatcgac ctgaccccac
     2041 tcagctcagg tcttggcaat ttgccgcggc ctttccagct gtgcctgcgc ccgcgctaag
     2101 aattctgcag tcgacggtac cgcgggcccg ggatccgccc ctctccctcc ccccccccta
     2161 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     2221 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     2281 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     2341 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     2401 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     2461 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     2521 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     2581 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     2641 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     2701 cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt caccggggtg
     2761 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2821 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2881 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2941 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3001 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3061 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3121 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3181 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3241 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3301 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3361 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3421 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     3481 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3541 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3601 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3661 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3721 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3781 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3841 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3901 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3961 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4021 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4081 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4141 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4201 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4261 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4321 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4441 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4501 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4561 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4621 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4681 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4741 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4801 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4861 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4921 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4981 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5041 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5101 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5161 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5221 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5281 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5341 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5401 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5461 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5521 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5581 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5641 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5701 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5761 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5821 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5881 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
