LOCUS       V020033                 8018 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020033
VERSION     V020033
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             625..3891
                     /label="Pdgfra(NM_001083316)"
                     /note="Pdgfra(NM_001083316)"
                     /gene="Pdgfra"
     CDS             3964..4683
                     /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    4806..4927
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(4934..5389)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5416..5520
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        5522..5879
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5914..6708
                     /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    6940..6987
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      7316..7904
                     /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 atctcgaggc caccatgggg acctcccacc aggtctttct ggtcctcagc
      661 tgtctcctca cagggccggg cctcatctcc tgccagctct tattaccctc tatcctccca
      721 aacgagaatg agaagattgt gcagctgaat tcgtcctttt ctctgagatg cgttggggag
      781 agtgaagtga gctggcagca ccccatgtct gaagaagacg accccaacgt ggaaatcaga
      841 agtgaggaga acaacagtgg cctctttgtc acagtgctgg aagtggttaa cgcctccgca
      901 gcccacaccg gatggtacac ttgctactac aaccacactc agacggatga gagtgagatc
      961 gaaggcaggc acatttacat ctatgtacca gacccagaca tggcctttgt gcctctcggg
     1021 atgaccgatt ccttagtcat cgtggaagag gatgactctg ccatcatacc ttgccgcacc
     1081 acagatccgg agactcaagt aaccttgcac aataacggga ggctggtgcc tgcctcctat
     1141 gacagcaggc agggcttcaa cggaaccttc agcgtggggc cttacatctg tgaggccacc
     1201 gtcaaaggga ggacgttcaa gaccagcgag tttaatgttt atgccttgaa agcaacgtca
     1261 gaactgaatc tggagatgga cgcacgccag actgtgtata aggcaggaga aacgatcgtg
     1321 gtgacctgtg ccgtctttaa caacgaggtc gttgacctgc agtggactta ccctggagaa
     1381 gtgagaaaca aaggtatcac catgctggag gagatcaaac tcccatccat caaactggtg
     1441 tacactttga ccgtccccaa ggccacggtg aaggacagtg gagagtatga atgtgctgcc
     1501 cgtcaggcca ctaaagaggt caaggaaatg aagagagtca ccatttctgt ccacgagaaa
     1561 ggcttcgtcg agattgagcc cacctttggc cagctggaag ctgtcaactt gcacgaagtc
     1621 agagagttcg tggtggaggt gcaggcctac ccgacgccca ggatatcgtg gctgaaggac
     1681 aacttgactc tgatagagaa tctcaccgag atcaccactg atgtacagaa gagtcaggaa
     1741 acaaggtatc agagcaaatt aaagctgatc cgggctaagg aagaagacag tggccattac
     1801 accattatag ttcaaaatga ggatgacgtg aagagttaca cgtttgagct gtcaacccta
     1861 gttcctgcat ccattttgga tctcgtagat gaccaccatg gctctggcgg gggacagact
     1921 gtgaggtgta cagccgaagg cacccctctt ccagaaatcg actggatgat ctgcaagcat
     1981 attaagaaat gtaataatga cacttcgtgg acagttttgg ccagcaatgt ctcaaatatt
     2041 atcacagagc tccctcgccg tggcaggagt accgtggagg gacgagtgtc cttcgccaaa
     2101 gtggaagaga ccatcgcagt tcgatgcctg gcaaagaaca acctcagcgt tgtggcccgt
     2161 gagctgaagc tggtggctcc cactctgcga tctgaactca cagtggcggc cgcagtgttg
     2221 gtgctgttgg tgattgtcat tgtctctctc atcgtcctgg tggtcatttg gaagcagaaa
     2281 ccacggtatg aaattcgctg gagggttatc gagtcaatca gccccgatgg acatgagtat
     2341 atctatgtgg accccatgca gttgccttac gactccagat gggaattccc gagagatggc
     2401 ctcgtgcttg gtcggatttt gggatccggt gcatttggga aagtggtcga aggtacagct
     2461 tatggattaa gccggtccca acctgtaatg aaggtggctg tgaagatgct caaacccaca
     2521 gccaggtcta gtgaaaagca ggctctcatg tctgagctga agataatgac tcacctgggg
     2581 ccacatttga acattgtgaa cttgctggga gcctgcacca agtcaggtcc catttacatc
     2641 atcacagaat actgcttcta tggggatttg gtcaactact tgcataagaa cagagatagc
     2701 ttcatgagcc aacacccaga gaagccgaag aaagacctgg acatctttgg attgaatcct
     2761 gcagacgaga gcacaagaag ttatgtgatt ttgtcttttg aaaataacgg cgactacatg
     2821 gacatgaagc aagctgatac cacacagtat gtccccatgc ttgaaaggaa agaggtttct
     2881 aaatactctg acatccagag atcgctgtac gatcggccag cctcctacaa gaagaaatcc
     2941 atgctagact cagaagtcaa aaacctcctt tcggacgatg actccgaggg tctgactttg
     3001 ctggatctat tgagcttcac ctatcaagtt gctcgaggaa tggaattttt ggcttcgaaa
     3061 aattgtgtcc accgggacct ggctgcccgc aacgtcctcc tggcgcaagg aaaaattgtg
     3121 aagatctgtg attttggcct ggccagagac atcatgcacg attccaacta cgtgtcaaaa
     3181 ggcagtactt tcctgcctgt gaagtggatg gcacccgaga gcatcttcga caacctctac
     3241 accacgctga gtgacgtctg gtcctatggc attctgctct gggaaatctt ttcccttggt
     3301 ggcacaccct atcctggcat gatggtcgat tctactttct acaataagat caagagtgga
     3361 taccggatgg ccaaacctga ccatgccacc agtgaagtct atgagatcat ggtgcagtgc
     3421 tggaacagtg agcccgagaa gagaccctcc ttctaccacc tcagcgagat agtggagaac
     3481 ctgttgccgg gacaatacaa gaagagttat gaaaagattc acctggactt cctaaagagt
     3541 gaccatccgg ccgtggcacg catgcgggtg gactctgata atgcgtacat cggtgtcact
     3601 tacaaaaatg aagaggataa gctgaaggac tgggaaggtg gcctggacga acagagactg
     3661 agcgctgaca gtggctacat catccccctg ccagacattg accctgttcc agaggaggaa
     3721 gacctgggca agaggaacag acacagctca cagacttcgg aagagagtgc catcgagaca
     3781 ggttccagta gttccacctt catcaagaga gaggacgaga ccatcgagga catcgacatg
     3841 atggatgaca ttggtataga ttcctcggac ctggtggagg acagcttcct gggtaccggc
     3901 ggcggaggga gtggaggagg gggaagcgga ggcggaggga gtcaggatcc accggtcgcc
     3961 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4021 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4081 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4141 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4201 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4261 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4321 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4381 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4441 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4501 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4561 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4621 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     4681 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     4741 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4801 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4861 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4921 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4981 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5041 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5101 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5161 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5221 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5281 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5341 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5401 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5461 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5521 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5581 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5641 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5701 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5761 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5821 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     5881 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5941 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6001 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6061 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     6121 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6181 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6241 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6301 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6361 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6421 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     6481 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6541 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6601 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6661 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6721 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6781 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6841 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     6901 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     6961 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7021 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7081 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     7141 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     7201 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7261 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7321 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7381 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7441 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     7501 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7561 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7621 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7681 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7741 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7801 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7861 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7921 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7981 cccctgattc tgtggataac cgtattaccg ccatgcat
//
