LOCUS       V024255                 6858 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024255
VERSION     V024255
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1055..1771
                     /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"
     CDS             1808..3106
                     /label="GSK3B(NM_002093)"
                     /note="GSK3B(NM_002093)"
                     /gene="GSK3B"
     primer_bind     complement(3115..3131)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3142..3764
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3913..4501)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4830..4877
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5109..5903)
                     /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"
     promoter        complement(5938..6295)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6297..6401)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6428..6856
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgcggc gcgccagatc tcaagcttgc caccatggtg agcaagggcg aggagctgtt
     1081 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1141 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1201 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1261 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1321 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1381 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1441 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1501 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1561 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1621 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1681 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1741 gatcactctc ggcatggacg agctgtacaa gggcggtgga ggcagtggtg gaggcggtag
     1801 ctctagaatg tcagggcggc ccagaaccac ctcctttgcg gagagctgca agccggtgca
     1861 gcagccttca gcttttggca gcatgaaagt tagcagagac aaggacggca gcaaggtgac
     1921 aacagtggtg gcaactcctg ggcagggtcc agacaggcca caagaagtca gctatacaga
     1981 cactaaagtg attggaaatg gatcatttgg tgtggtatat caagccaaac tttgtgattc
     2041 aggagaactg gtcgccatca agaaagtatt gcaggacaag agatttaaga atcgagagct
     2101 ccagatcatg agaaagctag atcactgtaa catagtccga ttgcgttatt tcttctactc
     2161 cagtggtgag aagaaagatg aggtctatct taatctggtg ctggactatg ttccggaaac
     2221 agtatacaga gttgccagac actatagtcg agccaaacag acgctccctg tgatttatgt
     2281 caagttgtat atgtatcagc tgttccgaag tttagcctat atccattcct ttggaatctg
     2341 ccatcgggat attaaaccgc agaacctctt gttggatcct gatactgctg tattaaaact
     2401 ctgtgacttt ggaagtgcaa agcagctggt ccgaggagaa cccaatgttt cgtatatctg
     2461 ttctcggtac tatagggcac cagagttgat ctttggagcc actgattata cctctagtat
     2521 agatgtatgg tctgctggct gtgtgttggc tgagctgtta ctaggacaac caatatttcc
     2581 aggggatagt ggtgtggatc agttggtaga aataatcaag gtcctgggaa ctccaacaag
     2641 ggagcaaatc agagaaatga acccaaacta cacagaattt aaattccctc aaattaaggc
     2701 acatccttgg actaaggatt cgtcaggaac aggacatttc acctcaggag tgcgggtctt
     2761 ccgaccccga actccaccgg aggcaattgc actgtgtagc cgtctgctgg agtatacacc
     2821 aactgcccga ctaacaccac tggaagcttg tgcacattca ttttttgatg aattacggga
     2881 cccaaatgtc aaactaccaa atgggcgaga cacacctgca ctcttcaact tcaccactca
     2941 agaactgtca agtaatccac ctctggctac catccttatt cctcctcatg ctcggattca
     3001 agcagctgct tcaaccccca caaatgccac agcagcgtca gatgctaata ctggagaccg
     3061 tggacagacc aataatgctg cttctgcatc agcttccaac tccacctgac cgcggtcata
     3121 gctgtttcct gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     3181 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     3241 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     3301 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     3361 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     3421 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3481 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3541 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3601 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     3661 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     3721 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg
     3781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4321 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4621 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     4681 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     4741 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     4801 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     4861 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     4921 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     4981 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     5041 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     5101 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     5161 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     5221 atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag
     5281 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     5341 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     5401 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     5461 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     5521 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     5581 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     5641 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     5701 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     5761 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     5821 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     5881 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     5941 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     6001 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     6061 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     6121 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6181 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     6241 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     6301 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6361 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6421 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6481 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6541 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6601 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6661 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6721 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6781 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6841 aaatttaacg cgaatttt
//
