LOCUS       V036009                 8454 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036009
VERSION     V036009
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             1366..5109
                     /label="APAF1(NM_181861)"
                     /note="APAF1(NM_181861)"
                     /gene="APAF1"
     polyA_signal    5242..5363
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(5370..5825)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5852..5956
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        5958..6315
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6350..7144
                     /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    7376..7423
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      7752..8340
                     /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 ctgtacaagg gcggcggagg gagtggagga gggggaagcc tcgagatgga tgcaaaagct
     1381 cgaaattgtt tgcttcaaca tagagaagct ctggaaaagg acatcaagac atcctacatc
     1441 atggatcaca tgattagtga tggattttta acaatatcag aagaggaaaa agtaagaaat
     1501 gagcccactc aacagcaaag agcagctatg ctgattaaaa tgatacttaa aaaagataat
     1561 gattcctacg tatcattcta caatgctcta ctacatgaag gatataaaga tcttgctgcc
     1621 cttctccatg atggcattcc tgttgtctct tcttccagtg gtaaagattc agttagtgga
     1681 ataacttcgt atgtaaggac agtcctgtgt gaaggtggag taccacagag gccagttgtt
     1741 tttgtcacaa ggaagaagct ggtgaatgca attcagcaga agctctccaa attgaaaggt
     1801 gaaccaggat gggtcaccat acatggaatg gcaggctgtg ggaagtctgt attagctgca
     1861 gaagctgtta gagatcattc ccttttagaa ggttgtttcc cagggggagt gcattgggtt
     1921 tcagttggga aacaagacaa atctgggctt ctgatgaaac tgcagaatct ttgcacacgg
     1981 ttggatcagg atgagagttt ttcccagagg cttccactta atattgaaga ggctaaagac
     2041 cgtctccgca ttctgatgct tcgcaaacac ccaaggtctc tcttgatctt ggatgatgtt
     2101 tgggactctt gggtgttgaa agcttttgac agtcagtgtc agattcttct tacaaccaga
     2161 gacaagagtg ttacagattc agtaatgggt cctaaatatg tagtccctgt ggagagttcc
     2221 ttaggaaagg aaaaaggact tgaaatttta tccctttttg ttaatatgaa gaaggcagat
     2281 ttgccagaac aagctcatag tattataaaa gaatgtaaag gctctcccct tgtagtatct
     2341 ttaattggtg cacttttacg tgattttccc aatcgctggg agtactacct caaacagctt
     2401 cagaataagc agtttaagag aataaggaaa tcttcgtctt atgattatga ggctctagat
     2461 gaagccatgt ctataagtgt tgaaatgctc agagaagaca tcaaagatta ttacacagat
     2521 ctttccatcc ttcagaagga cgttaaggtg cctacaaagg tgttatgtat tctctgggac
     2581 atggaaactg aagaagttga agacatactg caggagtttg taaataagtc tcttttattc
     2641 tgtgatcgga atggaaagtc gtttcgttat tatttacatg atcttcaagt agattttctt
     2701 acagagaaga attgcagcca gcttcaggat ctacataaga agataatcac tcagtttcag
     2761 agatatcacc agccgcatac tctttcacca gatcaggaag actgtatgta ttggtacaac
     2821 tttctggcct atcacatggc cagtgccaag atgcacaagg aactttgtgc tttaatgttt
     2881 tccctggatt ggattaaagc aaaaacagaa cttgtaggcc ctgctcatct gattcatgaa
     2941 tttgtggaat acagacatat actagatgaa aaggattgtg cagtcagtga gaattttcag
     3001 gagtttttat ctttaaatgg acaccttctt ggacgacagc catttcctaa tattgtacaa
     3061 ctgggtctct gtgagccgga aacttcagaa gtttatcagc aagctaagct gcaggccaag
     3121 caggaggtcg ataatggaat gctttacctg gaatggataa acaaaaaaaa catcacgaat
     3181 ctttcccgct tagttgtccg cccccacaca gatgctgttt accatgcctg cttttctgag
     3241 gatggtcaga gaatagcttc ttgtggagct gataaaacct tacaggtgtt caaagctgaa
     3301 acaggagaga aacttctaga aatcaaggct catgaggatg aagtgctttg ttgtgcattc
     3361 tctacagatg acagatttat agcaacctgc tcagtggata aaaaagtgaa gatttggaat
     3421 tctatgactg gggaactagt acacacctat gatgagcact cagagcaagt caattgctgc
     3481 catttcacca acagtagtca tcatcttctc ttagccactg ggtcaagtga ctgcttcctc
     3541 aaactttggg atttgaatca aaaagaatgt cgaaatacca tgtttggtca tacaaattca
     3601 gtcaatcact gcagattttc accagatgat aagcttttgg ctagttgttc agctgatgga
     3661 accttaaagc tttgggatgc gacatcagca aatgagagga aaagcattaa tgtgaaacag
     3721 ttcttcctaa atttggagga ccctcaagag gatatggaag tgatagtgaa gtgttgttcg
     3781 tggtctgctg atggtgcaag gataatggtg gcagcaaaaa ataaaatctt tctttttgac
     3841 attcatacta gtggcctatt gggagaaatc cacacgggcc atcacagcac catccagtac
     3901 tgtgacttct ccccacaaaa ccatttggca gtggttgctt tgtcccagta ctgtgtagag
     3961 ttgtggaata cagactcacg ttcaaaggtg gctgattgca gaggacattt aagttgggtt
     4021 catggtgtga tgttttctcc tgatggatca tcatttttga catcttctga tgaccagaca
     4081 atcaggctct gggagacaaa gaaagtatgt aagaactctg ctgtaatgtt aaagcaagaa
     4141 gtagatgttg tgtttcaaga aaatgaagtg atggtccttg cagttgacca tataagacgt
     4201 ctgcaactca ttaatggaag aacaggtcag attgattatc tgactgaagc tcaagttagc
     4261 tgctgttgct taagtccaca tcttcagtac attgcatttg gagatgaaaa tggagccatt
     4321 gagattttag aacttgtaaa caatagaatc ttccagtcca ggtttcagca caagaaaact
     4381 gtatggcaca tccagttcac agccgatgag aagactctta tttcaagttc tgatgatgct
     4441 gaaattcagg tatggaattg gcaattggac aaatgtatct ttctacgagg ccatcaggaa
     4501 acagtgaaag actttagact cttgaaaaat tcaagactgc tttcttggtc atttgatgga
     4561 acagtgaagg tatggaatat tattactgga aataaagaaa aagactttgt ctgtcaccag
     4621 ggtacagtac tttcttgtga catttctcac gatgctacca agttttcatc tacctctgct
     4681 gacaagactg caaagatctg gagttttgat ctccttttgc cacttcatga attgaggggc
     4741 cacaacggct gtgtgcgctg ctctgccttc tctgtggaca gtaccctgct ggcaacggga
     4801 gatgacaatg gagaaatcag gatatggaat gtctcaaacg gtgagcttct tcatttgtgt
     4861 gctccgcttt cagaagaagg agctgctacc catggaggct gggtgactga cctttgcttt
     4921 tctccagatg gcaaaatgct tatctctgct ggaggatata ttaagtggtg gaacgttgtc
     4981 actggggaat cctcacagac cttctacaca aatggaacca atcttaagaa aatacacgtg
     5041 tcccctgact tcaaaacata tgtgactgtg gataatcttg gtattttata tattttacag
     5101 actttagaat aaggatccac cggatctaga taactgatca taatcagcca taccacattt
     5161 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     5221 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5341 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     5401 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5461 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5521 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5581 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5641 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5701 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5761 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5821 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5881 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5941 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     6001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6061 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     6121 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     6181 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6241 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6301 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6361 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6421 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6481 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6541 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6601 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6661 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6721 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6781 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6841 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     6901 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6961 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     7021 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     7081 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     7141 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     7201 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     7261 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     7321 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     7381 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     7441 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     7501 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     7561 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     7621 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7681 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7741 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7801 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7861 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     7921 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7981 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8041 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     8101 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     8161 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     8221 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     8281 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8341 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8401 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
