LOCUS       V029002                 6709 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029002
VERSION     V029002
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2229
                     /label="GSK3B(NM_002093)"
                     /note="GSK3B(NM_002093)"
                     /gene="GSK3B"
     polyA_signal    2276..2500
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2546..2974
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2988..3317
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3384..4178
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4352..4473
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4522..4538)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4546..4562
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4570..4600)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4615..4636
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4924..5509)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5680..6540)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6541..6645)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtcagggc ggcccagaac cacctccttt
      961 gcggagagct gcaagccggt gcagcagcct tcagcttttg gcagcatgaa agttagcaga
     1021 gacaaggacg gcagcaaggt gacaacagtg gtggcaactc ctgggcaggg tccagacagg
     1081 ccacaagaag tcagctatac agacactaaa gtgattggaa atggatcatt tggtgtggta
     1141 tatcaagcca aactttgtga ttcaggagaa ctggtcgcca tcaagaaagt attgcaggac
     1201 aagagattta agaatcgaga gctccagatc atgagaaagc tagatcactg taacatagtc
     1261 cgattgcgtt atttcttcta ctccagtggt gagaagaaag atgaggtcta tcttaatctg
     1321 gtgctggact atgttccgga aacagtatac agagttgcca gacactatag tcgagccaaa
     1381 cagacgctcc ctgtgattta tgtcaagttg tatatgtatc agctgttccg aagtttagcc
     1441 tatatccatt cctttggaat ctgccatcgg gatattaaac cgcagaacct cttgttggat
     1501 cctgatactg ctgtattaaa actctgtgac tttggaagtg caaagcagct ggtccgagga
     1561 gaacccaatg tttcgtatat ctgttctcgg tactataggg caccagagtt gatctttgga
     1621 gccactgatt atacctctag tatagatgta tggtctgctg gctgtgtgtt ggctgagctg
     1681 ttactaggac aaccaatatt tccaggggat agtggtgtgg atcagttggt agaaataatc
     1741 aaggtcctgg gaactccaac aagggagcaa atcagagaaa tgaacccaaa ctacacagaa
     1801 tttaaattcc ctcaaattaa ggcacatcct tggactaagg attcgtcagg aacaggacat
     1861 ttcacctcag gagtgcgggt cttccgaccc cgaactccac cggaggcaat tgcactgtgt
     1921 agccgtctgc tggagtatac accaactgcc cgactaacac cactggaagc ttgtgcacat
     1981 tcattttttg atgaattacg ggacccaaat gtcaaactac caaatgggcg agacacacct
     2041 gcactcttca acttcaccac tcaagaactg tcaagtaatc cacctctggc taccatcctt
     2101 attcctcctc atgctcggat tcaagcagct gcttcaaccc ccacaaatgc cacagcagcg
     2161 tcagatgcta atactggaga ccgtggacag accaataatg ctgcttctgc atcagcttcc
     2221 aactccacct gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2281 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2341 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2401 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2461 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2521 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2581 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2641 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2701 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2761 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2821 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2881 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2941 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3001 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3061 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3241 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3301 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3361 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3421 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3481 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3541 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3601 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3661 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3721 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3781 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3841 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3901 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3961 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4021 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4081 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4141 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4201 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4261 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4321 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4381 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4441 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4501 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4561 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4621 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4681 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5101 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5341 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5521 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5581 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5641 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5701 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5761 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5821 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5881 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5941 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6001 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6061 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6121 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6181 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6241 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6301 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6361 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6421 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6481 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6541 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6601 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6661 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
