LOCUS       V023382                 6399 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023382
VERSION     V023382
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      626..635
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             635..664
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             668..685
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             689..712
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             731..2323
                     /label="PKM(NM_002654)"
                     /note="PKM(NM_002654)"
                     /gene="PKM"
     CDS             2345..3064
                     /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    3187..3308
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3315..3770)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3797..3901
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        3903..4260
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4295..5089
                     /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    5321..5368
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5697..6285
                     /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 atctcgagac cggtcgccac catggaacaa aaactcatct cagaagagga
      661 tctgagccat catcatcatc atcatgcgga ttacaaggat gacgacgata agctcaagct
      721 tcgaattctg atgtcgaagc cccatagtga agccgggact gccttcattc agacccagca
      781 gctgcacgca gccatggctg acacattcct ggagcacatg tgccgcctgg acattgattc
      841 accacccatc acagcccgga acactggcat catctgtacc attggcccag cttcccgatc
      901 agtggagacg ttgaaggaga tgattaagtc tggaatgaat gtggctcgtc tgaacttctc
      961 tcatggaact catgagtacc atgcggagac catcaagaat gtgcgcacag ccacggaaag
     1021 ctttgcttct gaccccatcc tctaccggcc cgttgctgtg gctctagaca ctaaaggacc
     1081 tgagatccga actgggctca tcaagggcag cggcactgca gaggtggagc tgaagaaggg
     1141 agccactctc aaaatcacgc tggataacgc ctacatggaa aagtgtgacg agaacatcct
     1201 gtggctggac tacaagaaca tctgcaaggt ggtggaagtg ggcagcaaga tctacgtgga
     1261 tgatgggctt atttctctcc aggtgaagca gaaaggtgcc gacttcctgg tgacggaggt
     1321 ggaaaatggt ggctccttgg gcagcaagaa gggtgtgaac cttcctgggg ctgctgtgga
     1381 cttgcctgct gtgtcggaga aggacatcca ggatctgaag tttggggtcg agcaggatgt
     1441 tgatatggtg tttgcgtcat tcatccgcaa ggcatctgat gtccatgaag ttaggaaggt
     1501 cctgggagag aagggaaaga acatcaagat tatcagcaaa atcgagaatc atgagggggt
     1561 tcggaggttt gatgaaatcc tggaggccag tgatgggatc atggtggctc gtggtgatct
     1621 aggcattgag attcctgcag agaaggtctt ccttgctcag aagatgatga ttggacggtg
     1681 caaccgagct gggaagcctg tcatctgtgc tactcagatg ctggagagca tgatcaagaa
     1741 gccccgcccc actcgggctg aaggcagtga tgtggccaat gcagtcctgg atggagccga
     1801 ctgcatcatg ctgtctggag aaacagccaa aggggactat cctctggagg ctgtgcgcat
     1861 gcagcacctg attgcccgtg aggcagaggc tgccatctac cacttgcaat tatttgagga
     1921 actccgccgc ctggcgccca ttaccagcga ccccacagaa gccaccgccg tgggtgccgt
     1981 ggaggcctcc ttcaagtgct gcagtggggc cataatcgtc ctcaccaagt ctggcaggtc
     2041 tgctcaccag gtggccagat accgcccacg tgcccccatc attgctgtga cccggaatcc
     2101 ccagacagct cgtcaggccc acctgtaccg tggcatcttc cctgtgctgt gcaaggaccc
     2161 agtccaggag gcctgggctg aggacgtgga cctccgggtg aactttgcca tgaatgttgg
     2221 caaggcccga ggcttcttca agaagggaga tgtggtcatt gtgctgaccg gatggcgccc
     2281 tggctccggc ttcaccaaca ccatgcgtgt tgttcctgtg ccgtcggatc caccggtcgc
     2341 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2401 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2461 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2521 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2581 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2641 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2701 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2761 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2821 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2881 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2941 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3001 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3061 gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     3121 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3181 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3241 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3301 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3361 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3421 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3481 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3541 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3601 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3661 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3721 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3781 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3841 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3901 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4141 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4201 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4261 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4321 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4381 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4441 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4501 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4561 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4621 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4681 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4741 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4801 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4861 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4921 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4981 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5041 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5101 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5161 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5221 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5281 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5341 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5401 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5461 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5521 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5581 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5641 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5701 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5761 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5821 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5881 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5941 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6001 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6061 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6121 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6181 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6241 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6301 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6361 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
