LOCUS       V025835                 6938 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025835
VERSION     V025835
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..2255
                     /label="CYP2U1(NM_183075)"
                     /note="CYP2U1(NM_183075)"
                     /gene="CYP2U1"
     misc_feature    2297..2883
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             2884..3603
                     /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    3726..3847
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3854..4309)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4336..4440
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4442..4799
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4834..5628
                     /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    5860..5907
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6236..6824
                     /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 accatgtcgt ctccggggcc gtcgcagccg ccggccgagg
      661 acccgccctg gcccgcgcgc ctcctgcgtg cgcctctggg gctgctgcgg ctggacccca
      721 gcgggggcgc gctgctgcta tgcggcctcg tagcgctgct gggctggagc tggctgcgga
      781 ggcgccgggc gcggggcatc ccgcccgggc ccacgccctg gcctctggtg ggcaacttcg
      841 gtcacgtgct gctgcctccc ttcctccggc ggcggagctg gctgagcagc aggaccaggg
      901 ccgcagggat tgatccctcg gtcataggcc cgcaggtgct cctggctcac ctagcccgcg
      961 tgtacggcag catcttcagc ttctttatcg gccactacct ggtggtggtc ctcagcgact
     1021 tccacagcgt gcgcgaggcg ctggtgcagc aggccgaggt cttcagcgac cgcccgcggg
     1081 tgccgctcat ctccatcgtg accaaggaga agggggttgt gtttgcacat tatggtcccg
     1141 tctggagaca acaaaggaag ttctctcatt caactcttcg tcattttggg ttgggaaaac
     1201 ttagcttgga gcccaagatt attgaggagt tcaaatatgt gaaagcagaa atgcaaaagc
     1261 acggagaaga ccccttctgc cctttctcca tcatcagcaa tgccgtctct aacatcattt
     1321 gctccttgtg ctttggccag cgctttgatt acactaatag tgagttcaag aaaatgcttg
     1381 gttttatgtc acgaggccta gaaatctgtc tgaacagtca agtcctcctg gtcaacatat
     1441 gcccttggct ttattacctt ccctttggac catttaagga attaagacaa attgaaaagg
     1501 atataaccag tttccttaaa aaaatcatca aagaccatca agagtctctg gatagagaga
     1561 accctcagga cttcatagac atgtaccttc tccacatgga agaggagagg aaaaataata
     1621 gtaacagcag ttttgatgaa gagtacttat tttatatcat tggggatctc tttattgctg
     1681 ggactgatac cacaactaac tctttgctct ggtgcctgct gtatatgtcg ctgaaccccg
     1741 atgtacaaga aaaggttcat gaagaaattg aaagagtcat tggcgccaac cgagctcctt
     1801 ccctcacaga caaggcccag atgccctaca cagaagccac catcatggaa gtgcagaggc
     1861 taactgtggt ggtgccgctt gccattcctc atatgacctc agagaacaca gtgctccaag
     1921 ggtataccat tcctaaaggc acattgatct tacccaacct gtggtcagta catagagacc
     1981 cagccatttg ggagaaaccg gaggatttct accctaatcg atttctggat gaccaaggac
     2041 aactaattaa aaaagaaacc tttattcctt ttgggatagg gaagcgggtg tgtatgggag
     2101 aacaactggc aaagatggaa ttattcctaa tgtttgtgag cctaatgcag agtttcgcat
     2161 ttgctttacc tgaggattct aagaagcccc tcctgactgg aagatttggt ctaactttag
     2221 ccccacatcc atttaatata actatttcaa ggagatgaga attctgcagt cgacggtacc
     2281 gcgggcccgg gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     2341 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     2401 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     2461 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     2521 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     2581 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     2641 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     2701 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     2761 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     2821 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     2881 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2941 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3001 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3061 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     3121 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3181 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3241 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3301 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     3361 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3421 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3481 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3541 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     3601 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3661 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3721 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3781 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3841 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3901 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3961 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4021 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4081 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4141 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4201 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4261 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     4321 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4381 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4441 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4501 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4561 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4621 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4681 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4741 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4801 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4861 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4921 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4981 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     5041 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5101 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5161 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5221 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5281 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5341 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     5401 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5461 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5521 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5581 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5641 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5701 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5761 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5821 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5881 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5941 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     6001 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     6061 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6121 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6181 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6241 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6301 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6361 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6421 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6481 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6541 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6601 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6661 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6721 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6781 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6841 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6901 cccctgattc tgtggataac cgtattaccg ccatgcat
//
