LOCUS       V022589                 6986 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022589
VERSION     V022589
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="enhanced YFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1360..3627
                     /label="Plcd1(NM_019676)"
                     /note="Plcd1(NM_019676)"
                     /gene="Plcd1"
     polyA_signal    3774..3895
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3902..4357)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4384..4488
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4490..4847
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4882..5676
                     /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    5908..5955
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6284..6872
                     /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 caccttcggc tacggcctgc agtgcttcgc 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 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga tggactccgg tcgggacttc
     1381 ctgaccttgc acgggctgca ggatgacccg gaccttcagg ccctcctgaa gggcagccag
     1441 cttctgaagg tgaagtccag ctcgtggcgt agagaacgct tctacaagct acaggaggac
     1501 tgcaagacca tctggcagga atctcgcaag gtcatgaggt ccccggagtc gcagctgttc
     1561 tccatcgagg acattcagga ggtgcggatg ggacaccgca cagaaggcct ggagaaattt
     1621 gccagagaca tacccgagga ccgatgcttc tccatcgtct tcaaggacca gcgcaatacc
     1681 ctagacctca tcgccccgtc tccagctgac gtccagcatt gggtgcaggg cctgcgcaag
     1741 atcatcgacc gctcgggctc catggaccag cggcagaagc tgcagcactg gattcactcc
     1801 tgcttgcgaa aggctgataa aaacaaggat aacaagatga acttcaagga ggtgaaggac
     1861 ttcctgaagg agctcaacgt acaggtggat gacagctacg ccaggaagat cttcagggaa
     1921 tgtgaccact cccagacaga ctcgctggag gatgaggaga ttgagacctt ctacaggatg
     1981 ctgacccagc gggcagagat cgaccgtgcc tttgcagagg ccgcaggttc tgcggagacc
     2041 ctgtcggtgg agaagctagt gacgttcctg cagcatcagc agcgggagga ggaggcgggg
     2101 ccagccttgg ccctctctct cattgagcgc tacgagccca gtgagactgc caaagcccag
     2161 cggcagatga ccaaggatgg cttcctcatg tacctgctgt ctgctgacgg taacgccttc
     2221 agcctggcgc accgccgtgt ctaccaggac atgaaccagc cgctcagcca ctacttagtg
     2281 tcttcttccc acaacaccta cctactggag gaccagctca cagggcccag cagcacggag
     2341 gcctacatcc gggctctgtg caaaggctgc cggtgtctgg agctggactg ctgggacggc
     2401 cccaaccagg aacccatcat ctaccatggc tacactttta cctctaagat acttttctgc
     2461 gacgtgctca gggccatccg ggactacgcc ttcaaggcat ccccctaccc agttatcctg
     2521 tccctggaga accactgcag cctggagcaa caacgagtca tggctcatca cctgagggcc
     2581 atcctggggc ccatgctgtt ggaccagccg ctggatgggg ttaccacgag cctgccttcg
     2641 cctgagcaac tgaaggaaaa gattctgttg aaagggaaga aactgggcgg gctgctacct
     2701 gctggagggg aaaacgggcc tgaagccacc gatgtgtctg acgaagatga ggctgctgaa
     2761 atggaagatg aggccgtgcg cagccaagtg caacacaagc ccaaggagga taagctaaag
     2821 ctggtgccgg agctctccga catggtcatt tactgcaaga gtgtccactt tggtggcttc
     2881 tccagtccta gcacctctgg gcaggcattc tatgagatgg cttccttctc cgagagccgt
     2941 gccctgcggc tgctgcaaga atcaggaaat agttttgtcc gccataacgt gggccatctg
     3001 agcaggatct acccagctgg gtggaggaca gattcctcca actacagtcc cgtggagatg
     3061 tggaatggag gctgccagat tgtggctctg aacttccaaa cccctgggcc agaaatggat
     3121 gtttaccttg gctgcttcca ggacaatgga ggttgtgggt atgtgctgaa accggccttc
     3181 ctgagagacc ctgacaccac cttcaactca cgggccctga ctcaggggcc ctggtgggcc
     3241 ccgaagaagc tccgagtctg gatcatctct ggacagcagc tgccgaaggt caataagaat
     3301 aagaattcta tcgtggaccc caaggtgata gtggagatcc acggtgtggg ccaggatgtg
     3361 gccagccgtc agacagcagt tattaccaat aatggtttca acccacggtg ggacacggag
     3421 tttgagtttg tggtagcagt gcctgacctt gcccttgtac gtttcatggt agaggattat
     3481 gactcctcct ctaagaacga ctttatcggc cagagtacta tcccctggaa cagcctcaag
     3541 caggggtacc gccatgtcca cctcttgtct aagaatggag acctgcatcc atcagccact
     3601 ctctttgtga agatctccat ccaggactag gtaccgcggg cccgggatcc accggatcta
     3661 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3721 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3781 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3841 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3901 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3961 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4021 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4081 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4141 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4201 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4261 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4321 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     4381 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4441 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     4501 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4561 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4621 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4681 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4741 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4801 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4861 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4921 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4981 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5041 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5101 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5161 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5221 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5281 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5341 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5401 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5461 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5521 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5581 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5641 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5701 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5761 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5821 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5881 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5941 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     6001 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     6061 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     6121 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6301 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6361 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6421 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6481 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6541 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6601 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6661 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6721 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6781 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6841 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6901 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6961 tggataaccg tattaccgcc atgcat
//
