LOCUS       V021322                 8378 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021322
VERSION     V021322
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..3025
                     /label="Ctnnb1(NM_007614)"
                     /note="Ctnnb1(NM_007614)"
                     /gene="Ctnnb1"
     CDS             3065..3130
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3154..3742
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3808..3856
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4058..4486
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4500..4829
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4896..5690
                     /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    5866..5987
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6036..6052)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6060..6076
                     /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(6084..6114)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6129..6150
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6438..7026)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7197..8057)
                     /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(8058..8162)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggctac tcaagctgac ctgatggagt tggacatggc
      721 catggagccg gacagaaaag ctgctgtcag ccactggcag cagcagtctt acttggattc
      781 tggaatccat tctggtgcca ccaccacagc tccttccctg agtggcaagg gcaaccctga
      841 ggaagaagat gttgacacct cccaagtcct ttatgaatgg gagcaaggct tttcccagtc
      901 cttcacgcaa gagcaagtag ctgatattga cgggcagtat gcaatgacta gggctcagag
      961 ggtccgagct gccatgttcc ctgagacgct agatgagggc atgcagatcc catccacgca
     1021 gtttgacgct gctcatccca ctaatgtcca gcgcttggct gaaccatcac agatgttgaa
     1081 acatgcagtt gtcaatttga ttaactatca ggatgacgcg gaacttgcca cacgtgcaat
     1141 tcctgagctg acaaaactgc taaacgatga ggaccaggtg gtagttaata aagctgctgt
     1201 tatggtccat cagctttcca aaaaggaagc ttccagacat gccatcatgc gctcccctca
     1261 gatggtgtct gccattgtac gcaccatgca gaatacaaat gatgtagaga cagctcgttg
     1321 tactgctggg actctgcaca acctttctca ccaccgcgag ggcttgctgg ccatctttaa
     1381 gtctggtggc atcccagcgc tggtgaaaat gcttgggtca ccagtggatt ctgtactgtt
     1441 ctacgccatc acgacactgc ataatctcct gctccatcag gaaggagcta aaatggcagt
     1501 gcgcctagct ggtggactgc agaaaatggt tgctttgctc aacaaaacaa acgtgaaatt
     1561 cttggctatt acaacagact gccttcagat cttagcttat ggcaatcaag agagcaagct
     1621 catcattctg gccagtggtg gaccccaagc cttagtaaac ataatgagga cctacactta
     1681 tgagaagctt ctgtggacca caagcagagt gctgaaggtg ctgtctgtct gctctagcaa
     1741 caagccggcc attgtagaag ctggtgggat gcaggcactg gggcttcatc tgacagaccc
     1801 aagtcagcga cttgttcaaa acagtctttg gactctcaga aacctttcag atgcagcgac
     1861 taagcaggaa gggatggaag gcctccttgg gactctagtg cagcttctgg gttccgatga
     1921 tataaatgtg gtcacctgtg cagctggaat tctctctaac ctcacttgca ataattacaa
     1981 aaacaagatg atggtgtgcc aagtgggtgg catagaggct cttgtacgca ccgtccttcg
     2041 tgctggtgac agggaagaca tcactgagcc tgccatctgt gctcttcgtc atctgaccag
     2101 ccggcatcag gaagccgaga tggcccagaa tgccgttcgc cttcattatg gactgcctgt
     2161 tgtggttaaa ctcctgcacc caccatccca ctggcctctg ataaaggcaa ctgttggatt
     2221 gattcgaaac cttgcccttt gcccagcaaa tcatgcgcct ttgcgggaac agggtgctat
     2281 tccacgacta gttcagctgc ttgtacgagc acatcaggac acccaacggc gcacctccat
     2341 gggtggaacg cagcagcagt ttgtggaggg cgtgcgcatg gaggagatag tagaagggtg
     2401 tactggagct ctccacatcc ttgctcggga cgttcacaac cggattgtaa tccgaggact
     2461 caataccatt ccattgtttg tgcagttgct ttattctccc attgaaaata tccaaagagt
     2521 agctgcaggg gtcctctgtg aacttgctca ggacaaggag gctgcagagg ccattgaagc
     2581 tgagggagcc acagctcccc tgacagagtt actccactcc aggaatgaag gcgtggcaac
     2641 atacgcagct gctgtcctat tccgaatgtc tgaggacaag ccacaggatt acaagaagcg
     2701 gctttcagtc gagctgacca gttccctctt caggacagag ccaatggctt ggaatgagac
     2761 tgcagatctt ggactggaca ttggtgccca gggagaagcc cttggatatc gccaggatga
     2821 tcccagctac cgttcttttc actctggtgg atacggccag gatgccttgg ggatggaccc
     2881 tatgatggag catgagatgg gtggccacca ccctggtgct gactatccag ttgatgggct
     2941 gcctgatctg ggacacgccc aggacctcat ggatgggctg cccccaggtg atagcaatca
     3001 gctggcctgg tttgatactg acctggaatt ctgcagatat ccagcacagt ggcggccgct
     3061 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     3121 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     3181 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3241 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3301 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3361 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3421 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3481 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3541 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3601 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3661 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3721 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     3781 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     3841 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3901 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     3961 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4021 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4081 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4141 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4201 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4261 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4321 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4381 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     4441 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4501 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4561 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4621 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4681 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4741 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4801 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4861 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4921 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4981 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5041 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5101 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5161 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5221 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5281 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5341 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5401 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5461 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5521 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5581 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5641 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5701 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5761 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5821 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6001 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6601 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6841 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8161 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     8221 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     8281 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     8341 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
