LOCUS       V032292                 5836 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032292
VERSION     V032292
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"
     regulatory      609..618
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             618..683
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             687..1748
                     /label="OTC(NM_000531)"
                     /note="OTC(NM_000531)"
                     /gene="OTC"
     CDS             1782..2501
                     /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    2624..2745
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(2752..3207)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3234..3338
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3340..3697
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3732..4526
                     /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    4758..4805
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5134..5722
                     /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 gctagcgttt
      601 aaacttaagc caccatggac tacaaagacc atgacggtga ttataaagat catgacatcg
      661 actacaagga tgacgatgac aagcttatgc tgtttaatct gaggatcctg ttaaacaatg
      721 cagcttttag aaatggtcac aacttcatgg ttcgaaattt tcggtgtgga caaccactac
      781 aaaataaagt gcagctgaag ggccgtgacc ttctcactct aaaaaacttt accggagaag
      841 aaattaaata tatgctatgg ctatcagcag atctgaaatt taggataaaa cagaaaggag
      901 agtatttgcc tttattgcaa gggaagtcct taggcatgat ttttgagaaa agaagtactc
      961 gaacaagatt gtctacagaa acaggctttg cacttctggg aggacatcct tgttttctta
     1021 ccacacaaga tattcatttg ggtgtgaatg aaagtctcac ggacacggcc cgtgtattgt
     1081 ctagcatggc agatgcagta ttggctcgag tgtataaaca atcagatttg gacaccctgg
     1141 ctaaagaagc atccatccca attatcaatg ggctgtcaga tttgtaccat cctatccaga
     1201 tcctggctga ttacctcacg ctccaggaac actatagctc tctgaaaggt cttaccctca
     1261 gctggatcgg ggatgggaac aatatcctgc actccatcat gatgagcgca gcgaaattcg
     1321 gaatgcacct tcaggcagct actccaaagg gttatgagcc ggatgctagt gtaaccaagt
     1381 tggcagagca gtatgccaaa gagaatggta ccaagctgtt gctgacaaat gatccattgg
     1441 aagcagcgca tggaggcaat gtattaatta cagacacttg gataagcatg ggacaagaag
     1501 aggagaagaa aaagcggctc caggctttcc aaggttacca ggttacaatg aagactgcta
     1561 aagttgctgc ctctgactgg acatttttac actgcttgcc cagaaagcca gaagaagtgg
     1621 atgatgaagt cttttattct cctcgatcac tagtgttccc agaggcagaa aacagaaagt
     1681 ggacaatcat ggctgtcatg gtgtccctgc tgacagatta ctcacctcag ctccagaagc
     1741 ctaaatttgg aggaggggga agcgatccac cggtcgccac catggtgagc aagggcgagg
     1801 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1861 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1921 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1981 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2041 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2101 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2161 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2221 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2281 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2341 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2401 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2461 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat
     2521 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2581 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2641 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2701 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2761 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2821 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2881 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2941 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3001 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3061 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3121 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3181 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3241 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3301 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3361 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3421 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3481 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3541 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3601 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3661 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3721 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3781 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3841 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3901 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3961 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4021 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4081 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4141 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4201 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4261 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4321 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4381 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4441 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4501 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4561 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4621 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4681 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4741 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4801 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4861 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4921 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4981 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5041 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5101 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5161 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5221 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5281 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5341 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5401 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5461 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5521 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5581 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5641 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5701 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5761 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5821 tattaccgcc atgcat
//
