LOCUS       sequence-227467-        7805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7805)
  AUTHORS   Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B, 
            Kinzler KW
  TITLE     Activation of beta-catenin-Tcf signaling in colon cancer by 
            mutations in beta-catenin or APC.
  JOURNAL   Science. 1997 Mar 21. 275(5307):1787-90.
  PUBMED    9065402
REFERENCE   2  (bases 1 to 7805)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            1997 Mar 21. 275(5307):1787-90."
COMMENT     Sequence Label: pCI-neo beta catenin S33Y
FEATURES             Location/Qualifiers
     source          1..7805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     589..613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          753..885
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     868..887
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        930..948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             960..3305
                     /codon_start=1
                     /gene="CTNNB1"
                     /product="protein involved both in cell-cell adhesion as 
                     part of the cadherin protein complex, and in gene 
                     transcription as part of the Wnt signaling pathway"
                     /label=CTNNB1
                     /note="also known as catenin beta-1; related to Drosophila 
                     armadillo"
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDYGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     polyA_signal    3368..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3379..3398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3433..3452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3675..4130
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3762..3781)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3972..3993
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4247..4604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4247..4438
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4270..4290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4455..4590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4655..5449
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4709..4728)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5319..5338
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5513..5561
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5602..5621)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5721..5743
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5781..5799)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5867..5971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5972..6040,6041..6832)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6190..6209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7003..7591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7492..7511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      601 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      661 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agtgactctc ttaaggtagc
      721 cttgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
      781 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
      841 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
      901 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agaaccacca
      961 tggctactca agctgatttg atggagttgg acatggccat ggaaccagac agaaaagcgg
     1021 ctgttagtca ctggcagcaa cagtcttacc tggactatgg aatccattct ggtgccacta
     1081 ccacagctcc ttctctgagt ggtaaaggca atcctgagga agaggatgtg gatacctccc
     1141 aagtcctgta tgagtgggaa cagggatttt ctcagtcctt cactcaagaa caagtagctg
     1201 atattgatgg acagtatgca atgactcgag ctcagagggt acgagctgct atgttccctg
     1261 agacattaga tgagggcatg cagatcccat ctacacagtt tgatgctgct catcccacta
     1321 atgtccagcg tttggctgaa ccatcacaga tgctgaaaca tgcagttgta aacttgatta
     1381 actatcaaga tgatgcagaa cttgccacac gtgcaatccc tgaactgaca aaactgctaa
     1441 atgacgagga ccaggtggtg gttaataagg ctgcagttat ggtccatcag ctttctaaaa
     1501 aggaagcttc cagacacgct atcatgcgtt ctcctcagat ggtgtctgct attgtacgta
     1561 ccatgcagaa tacaaatgat gtagaaacag ctcgttgtac cgctgggacc ttgcataacc
     1621 tttcccatca tcgtgagggc ttactggcca tctttaagtc tggaggcatt cctgccctgg
     1681 tgaaaatgct tggttcacca gtggattctg tgttgtttta tgccattaca actctccaca
     1741 accttttatt acatcaagaa ggagctaaaa tggcagtgcg tttagctggt gggctgcaga
     1801 aaatggttgc cttgctcaac aaaacaaatg ttaaattctt ggctattacg acagactgcc
     1861 ttcaaatttt agcttatggc aaccaagaaa gcaagctcat catactggct agtggtggac
     1921 cccaagcttt agtaaatata atgaggacct atacttacga aaaactactg tggaccacaa
     1981 gcagagtgct gaaggtgcta tctgtctgct ctagtaataa gccggctatt gtagaagctg
     2041 gtggaatgca agctttagga cttcacctga cagatccaag tcaacgtctt gttcagaact
     2101 gtctttggac tctcaggaat ctttcagatg ctgcaactaa acaggaaggg atggaaggtc
     2161 tccttgggac tcttgttcag cttctgggtt cagatgatat aaatgtggtc acctgtgcag
     2221 ctggaattct ttctaacctc acttgcaata attataagaa caagatgatg gtctgccaag
     2281 tgggtggtat agaggctctt gtgcgtactg tccttcgggc tggtgacagg gaagacatca
     2341 ctgagcctgc catctgtgct cttcgtcatc tgaccagccg acaccaagaa gcagagatgg
     2401 cccagaatgc agttcgcctt cactatggac taccagttgt ggttaagctc ttacacccac
     2461 catcccactg gcctctgata aaggctactg ttggattgat tcgaaatctt gccctttgtc
     2521 ccgcaaatca tgcacctttg cgtgagcagg gtgccattcc acgactagtt cagttgcttg
     2581 ttcgtgcaca tcaggatacc cagcgccgta cgtccatggg tgggacacag cagcaatttg
     2641 tggagggggt ccgcatggaa gaaatagttg aaggttgtac cggagccctt cacatcctag
     2701 ctcgggatgt tcacaaccga attgttatca gaggactaaa taccattcca ttgtttgtgc
     2761 agctgcttta ttctcccatt gaaaacatcc aaagagtagc tgcaggggtc ctctgtgaac
     2821 ttgctcagga caaggaagct gcagaagcta ttgaagctga gggagccaca gctcctctga
     2881 cagagttact tcactctagg aatgaaggtg tggcgacata tgcagctgct gttttgttcc
     2941 gaatgtctga ggacaagcca caagattaca agaaacggct ttcagttgag ctgaccagct
     3001 ctctcttcag aacagagcca atggcttgga atgagactgc tgatcttgga cttgatattg
     3061 gtgcccaggg agaacccctt ggatatcgcc aggatgatcc tagctatcgt tcttttcact
     3121 ctggtggata tggccaggat gccttgggta tggaccccat gatggaacat gagatgggtg
     3181 gccaccaccc tggtgctgac tatccagttg atgggctgcc agatctgggg catgcccagg
     3241 acctcatgga tgggctgcct ccaggtgaca gcaatcagct ggcctggttt gatactgacc
     3301 tgtaatctag agtcgacccg ggcggccgct tccctttagt gagggttaat gcttcgagca
     3361 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3421 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3481 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     3541 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatccgata aggatcgatc
     3601 cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     3661 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3721 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3781 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3841 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3901 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3961 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4021 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4081 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     4141 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac
     4201 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga
     4261 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4441 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4501 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4561 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg
     4621 acacaacagt ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc
     4681 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4741 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4801 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4861 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4921 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4981 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5041 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5101 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5161 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5221 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5281 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5341 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5401 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5461 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     5521 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     5581 ccgcgtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5641 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5701 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5761 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5821 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5881 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5941 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6001 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6061 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6121 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6181 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6241 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6301 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6361 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6421 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6481 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6541 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6601 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6661 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6721 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6781 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6841 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6901 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6961 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7021 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7081 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7141 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7201 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7261 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7321 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7381 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7441 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7501 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7561 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7621 cctggccttt tgctggcctt ttgctcacat ggctcgacag atcgatcttc aatattggcc
     7681 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
     7741 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     7801 atgtt
//
