LOCUS       V025693                11062 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025693
VERSION     V025693
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      629..638
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             638..703
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             710..6190
                     /label="MAP2(NM_002374)"
                     /note="MAP2(NM_002374)"
                     /gene="MAP2"
     polyA_signal    6231..6455
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     promoter        6471..6994
                     /label="mCMV promoter"
     CDS             7008..7727
                     /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    7850..7971
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(7978..8433)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        8460..8564
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        8566..8923
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             8958..9752
                     /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    9984..10031
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      10360..10948
                     /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 atctcgagct ccgaattcgc caccatggac tacaaagacc atgacggtga
      661 ttataaagat catgacatcg actacaagga tgacgatgac aagaagctta tggcagatga
      721 acggaaagat gaagcaaagg cacctcactg gacctcagca ccgctaacag aggcatctgc
      781 acactcacat ccacctgaga ttaaggatca aggcggagca ggggaaggac ttgtccgaag
      841 cgccaatgga ttcccataca gggaggatga agagggtgcc tttggagagc atgggtcaca
      901 gggcacctat tcaaatacca aagagaatgg gatcaacgga gagctgacct cagctgacag
      961 agaaacagca gaggaggtgt ctgcaaggat agttcaagta gtcactgctg aggctgtagc
     1021 agtcctgaaa ggtgaacaag agaaagaagc tcaacataaa gaccagactg cagctctgcc
     1081 tttagcagct gaagaaacag ctaatctgcc tccttctcca cccccatcac ctgcctcaga
     1141 acagactgtc acagtggagg aagatttact tacagcctcg aagatggagt tccacgatca
     1201 acaggaattg actccctcta cagctgagcc ttcagaccag aaggaaaagg agtcagagaa
     1261 gcaaagtaag cctggtgaag accttaaaca tgctgcctta gtttctcagc cagagacaac
     1321 taaaacttac cctgataaaa aggacatgca aggcacggaa gaagaaaaag cacccctagc
     1381 tttgtttggg cacactcttg ttgccagcct ggaagacatg aaacagaaga cagaaccaag
     1441 ccttgtagta cctggcattg acctccctaa agagcctcca actccaaaag aacaaaagga
     1501 ctggttcatc gaaatgccaa cggaagcaaa aaaggatgag tggggtttag ttgcccccat
     1561 atctcctggc cctctgactc ccatgaggga aaaagatgta tttgatgata tcccaaaatg
     1621 ggaagggaaa cagtttgatt ctcccatgcc aagtcccttt caagggggaa gcttcactct
     1681 tcctttagat gtcatgaaga atgaaatagt tacagaaaca tcgccctttg cccctgcctt
     1741 tttacagcca gatgacaaaa aatctctgca acaaaccagt ggcccagcta ctgccaaaga
     1801 tagttttaaa attgaagagc cccatgaggc taaacctgac aaaatggcag aagcaccacc
     1861 ctcagaggca atgaccttac ccaaagatgc tcacattcca gttgtagaag aacatgttat
     1921 ggggaaagtt ttagaggaag aaaaggaggc cataaatcaa gagactgtgc agcaaaggga
     1981 tactttcacc cccagtggac aggaacctat acttactgaa aaggaaactg agctgaagct
     2041 tgaagaaaaa accaccattt ctgacaaaga agctgtgcca aaagagagta aacccccaaa
     2101 acctgcagat gaagaaatag gcataattca gacctccaca gagcacactt tctcagaaca
     2161 gaaagaccaa gagcctacca cagatatgtt gaaacaggac tcgttccctg taagtttgga
     2221 gcaagcagtt acagattcag ccatgacctc taaaacactg gagaaagcca tgaccgaacc
     2281 atctgcatta attgaaaaga gctcaattca ggaacttttt gaaatgagag ttgatgacaa
     2341 agataagatt gaaggagttg gagctgcaac atcagctgag cttgatatgc cattttatga
     2401 agataaatca ggaatgtcca agtactttga aacatctgcc ttgaaagaag aagcaacaaa
     2461 aagcattgag ccaggcagtg attactatga actgagtgac actagagaaa gtgtccatga
     2521 gtctattgat accatgtctc ccatgcataa aaatggtgac aaggagtttc aaacaggaaa
     2581 agaatcccag cccagtcctc cagcacaaga agcagggtac agcactctcg cacagagtta
     2641 tccatcagat ttacctgaag aacccagttc tcctcaagaa agaatgttca ctattgatcc
     2701 aaaagtgtat ggagagaaaa gggacctcca cagtaagaat aaggatgatt tgacccttag
     2761 caggagttta ggacttggtg gtaggtctgc aatagaacaa agaagcatgt caatcaattt
     2821 gccgatgtct tgcctagatt ccatagccct tggatttaac tttggtcggg gacatgatct
     2881 ttctcctctg gcttccgata ttctaaccaa cactagtgga agtatggatg aaggggatga
     2941 ttaccttcca gccaccacac ctgcactgga gaaagcccct tgcttccctg tagaaagcaa
     3001 agaggaagaa cagatagaga aagtaaaagc tactggagaa gaaagtactc aagcggagat
     3061 atcatgtgag tctcctttcc tagccaaaga tttttacaaa aatggtactg tcatggcacc
     3121 tgaccttcct gaaatgctag atctggcagg cacaaggtca agattggctt ctgtgagtgc
     3181 agatgctgag gttgccagga ggaaatcagt cccatcagag actgtggttg aggatagtcg
     3241 tactggcttg cccccggtaa ctgatgaaaa ccatgtcatt gtaaaaacgg acagtcagct
     3301 cgaagacctg ggctactgtg tgttcaataa gtacacagtc ccattgccat cacctgttca
     3361 agacagtgag aatttatcag gggagagtgg taccttttac gaaggcactg atgataaagt
     3421 tcgaagagat ttggccacag acctttcact gattgaagtg aaactggcag cagccggaag
     3481 agtcaaagat gagttcagtg ttgacaaaga agcatccgcg catatctctg gtgacaaatc
     3541 aggactgagt aaggagtttg accaagagaa gaaagctaat gataggttgg atactgtact
     3601 agaaaagagt gaagaacatg ctgattcaaa agaacatgcc aagaaaactg aagaggctgg
     3661 tgatgaaata gaaacattcg gattaggagt aacctatgag caagctttgg ccaaagattt
     3721 gtcaatacca acagatgcat cctctgagaa agcagagaag ggtcttagtt cagtgccaga
     3781 gatagctgag gtagaaccat ccaaaaaggt ggaacaaggt ctggattttg ctgtccaggg
     3841 tcaactagat gttaaaatta gtgactttgg acagatggct tcagggctaa acatagatga
     3901 tagaagggca acagagctaa aacttgaggc tacacaggac atgaccccct catccaaagc
     3961 accgcaggag gcagatgcat ttatgggtgt tgagtctggc cacatgaaag aaggcactaa
     4021 agttagtgag acagaagtca aagagaaggt ggccaagcct gacttggtgc accaggaggc
     4081 tgtagacaag gaggagtcct atgaatctag tggtgagcat gaaagtctca ccatggagtc
     4141 cttgaaagct gatgagggca agaaggaaac atctccagaa tcatctctaa ttcaagatga
     4201 gattgccgtc aaattgtcag tggaaatacc ttgcccacct gctgtttcag aggctgattt
     4261 agccacagat gagagagctg atgtccagat ggaatttatt caggggccaa aagaagaaag
     4321 caaagagacc ccagatatat ccatcacgcc ttctgatgtt gcagagccat tgcatgaaac
     4381 gatcgtatct gaaccagcag agattcagag tgaggaagaa gagatagaag cccagggaga
     4441 atatgataaa ctgctcttcc gctcagacac ccttcagata actgacctgg gtgtctcagg
     4501 tgccagggag gaatttgtgg agacctgccc aagtgaacac aaaggagtga ttgagtctgt
     4561 tgtgaccatc gaggatgatt tcatcactgt agtgcaaacc acaactgatg aaggggagtc
     4621 agggtcccac agcgtgcgtt ttgcagccct agagcagcct gaggtggaaa ggagaccatc
     4681 tcctcatgat gaagaagagt ttgaagtaga agaggcagct gaagcccagg cagaacccaa
     4741 agatggttcc ccagaggctc cagcttcccc tgagagagaa gaggttgcac tttctgaata
     4801 taagacagaa acctatgacg attacaaaga tgagaccacc attgacgact ccatcatgga
     4861 cgctgacagc ctctgggtgg acactcaaga tgatgatagg agcatcatga cagaacagtt
     4921 agaaactatt cctaaagagg agaaagctga aaaggaagct cggagatcat ctcttgagaa
     4981 acatagaaaa gaaaagcctt ttaaaaccgg gagaggcaga atttccactc ctgaaagaaa
     5041 agtagctaaa aaggaaccta gcacagtctc cagagatgaa gtgagaagga aaaaagcagt
     5101 ttataagaag gctgaacttg ctaaaaaaac agaagttcag gcccactctc cctccaggaa
     5161 attcatttta aaacctgcta tcaaatatac tagaccaact catctctcct gtgttaagcg
     5221 gaaaaccaca gcagcaggtg gggaatcagc tctggctccc agtgtattta aacaggcaaa
     5281 ggacaaagtc tctgacggag taaccaagag cccagaaaag cgctcttctc tcccaagacc
     5341 ttcctccatt ctccctcctc ggcgaggtgt gtcaggagac agagatgaga attccttctc
     5401 tctcaacagt tctatctctt cttcagcacg gcggaccacc aggtcagagc caattcgcag
     5461 agcagggaag agtggtacct caacacccac tacccctggg tctactgcca tcactcctgg
     5521 caccccacca agttattctt cacgcacacc aggcactcct ggaaccccta gctatcccag
     5581 gacccctcac acaccaggaa cccccaagtc tgccatcttg gtgccgagtg agaagaaggt
     5641 cgccatcata cgtactcctc caaaatctcc tgcgactccc aagcagcttc ggcttattaa
     5701 ccaaccactg ccagacctga agaatgtcaa atccaaaatc ggatcaacag acaacatcaa
     5761 ataccagcct aaaggggggc aggtacaaat tgttaccaag aaaatagacc taagccatgt
     5821 gacatccaaa tgtggctctc tgaagaacat ccgccacagg ccaggtggcg gacgtgtgaa
     5881 aattgagagt gtaaaactag atttcaaaga aaaggcccaa gctaaagttg gttctcttga
     5941 taatgctcat catgtacctg gaggtggtaa tgtcaagatt gacagccaaa agttgaactt
     6001 cagagagcat gctaaagccc gtgtggacca tggggctgag atcattacac agtccccagg
     6061 cagatccagc gtggcatcac cccgacgact cagcaatgtc tcctcgtctg gaagcatcaa
     6121 cctgctcgaa tctcctcagc ttgccacttt ggctgaggat gtcactgctg cactcgctaa
     6181 gcagggcttg tgaggatccg ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     6241 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     6301 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6361 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     6421 tagcaggcat gctggggatg cggtgggctc tatggtctag agatatactg agtcattagg
     6481 gactttccaa tgggttttgc ccagtacata aggtcaatag gggtgaatca acaggaaagt
     6541 cccattggag ccaagtacac tgagtcaata gggactttcc attgggtttt gcccagtaca
     6601 aaaggtcaat agggggtgag tcaatgggtt tttcccatta ttggcacgta cataaggtca
     6661 ataggggtga gtcattgggt ttttccagcc aatttaatta aaacgccatg tactttccca
     6721 ccattgacgt caatgggcta ttgaaactaa tgcaacgtga cctttaaacg gtactttccc
     6781 atagctgatt aatgggaaag taccgttctc gagccaatac acgtcaatgg gaagtgaaag
     6841 ggcagccaaa acgtaacacc gccccggttt tcccctggaa attccatatt ggcacgcatt
     6901 ctattggctg agctgcgttc tacgtgggta taagaggcgc gaccagcgtc ggtaccgtcg
     6961 cagtcttcgg tctgaccacc gtagaacgca gatcaccggt cgccaccatg gtgagcaagg
     7021 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     7081 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     7141 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     7201 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     7261 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     7321 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     7381 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     7441 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     7501 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     7561 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     7621 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     7681 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     7741 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     7801 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     7861 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7921 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     7981 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     8041 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     8101 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     8161 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     8221 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     8281 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     8341 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     8401 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     8461 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8521 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     8581 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     8641 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     8701 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     8761 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     8821 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     8881 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     8941 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     9001 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     9061 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     9121 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     9181 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     9241 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     9301 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     9361 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     9421 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     9481 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     9541 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     9601 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     9661 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     9721 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     9781 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     9841 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     9901 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     9961 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
    10021 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
    10081 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
    10141 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
    10201 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
    10261 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    10321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    10381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    10441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    10501 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    10561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
    10621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
    10681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
    10741 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
    10801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
    10861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
    10921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
    10981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
    11041 taaccgtatt accgccatgc at
//
