LOCUS       sequence-183792-        7811 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7811)
  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 7811)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 WT
FEATURES             Location/Qualifiers
     source          1..7811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        422..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          794..926
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     909..928
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        971..989
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1007..3352
                     /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="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     polyA_signal    3415..3536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3480..3499
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3722..4177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3809..3828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4019..4040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4294..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4294..4485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4702..5496
                     /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(4756..4775)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5366..5385
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5560..5608
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5649..5668)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5768..5790
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5828..5846)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5914..6018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6019..6087,6088..6879)
                     /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(6237..6256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7050..7638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7539..7558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtacatttat attggctcat gtccaatatg accgccatgt tggcattgat tattgactag
       61 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      121 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      181 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      241 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      301 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      361 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      421 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
      481 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      541 ctttccaaaa tgtcgtaaca actgcgatcg cccgccccgt tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcactaga
      661 agctttattg cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca
      721 acagtctcga acttaagctg cagtgactct cttaaggtag ccttgcagaa gttggtcgtg
      781 aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
      841 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
      901 catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg
      961 ctagagtact taatacgact cactataggc tagcctcgac accaccatgg ctactcaagc
     1021 tgatttgatg gagttggaca tggccatgga accagacaga aaagcggctg ttagtcactg
     1081 gcagcaacag tcttacctgg actctggaat ccattctggt gccactacca cagctccttc
     1141 tctgagtggt aaaggcaatc ctgaggaaga ggatgtggat acctcccaag tcctgtatga
     1201 gtgggaacag ggattttctc agtccttcac tcaagaacaa gtagctgata ttgatggaca
     1261 gtatgcaatg actcgagctc agagggtacg agctgctatg ttccctgaga cattagatga
     1321 gggcatgcag atcccatcta cacagtttga tgctgctcat cccactaatg tccagcgttt
     1381 ggctgaacca tcacagatgc tgaaacatgc agttgtaaac ttgattaact atcaagatga
     1441 tgcagaactt gccacacgtg caatccctga actgacaaaa ctgctaaatg acgaggacca
     1501 ggtggtggtt aataaggctg cagttatggt ccatcagctt tctaaaaagg aagcttccag
     1561 acacgctatc atgcgttctc ctcagatggt gtctgctatt gtacgtacca tgcagaatac
     1621 aaatgatgta gaaacagctc gttgtaccgc tgggaccttg cataaccttt cccatcatcg
     1681 tgagggctta ctggccatct ttaagtctgg aggcattcct gccctggtga aaatgcttgg
     1741 ttcaccagtg gattctgtgt tgttttatgc cattacaact ctccacaacc ttttattaca
     1801 tcaagaagga gctaaaatgg cagtgcgttt agctggtggg ctgcagaaaa tggttgcctt
     1861 gctcaacaaa acaaatgtta aattcttggc tattacgaca gactgccttc aaattttagc
     1921 ttatggcaac caagaaagca agctcatcat actggctagt ggtggacccc aagctttagt
     1981 aaatataatg aggacctata cttacgaaaa actactgtgg accacaagca gagtgctgaa
     2041 ggtgctatct gtctgctcta gtaataagcc ggctattgta gaagctggtg gaatgcaagc
     2101 tttaggactt cacctgacag atccaagtca acgtcttgtt cagaactgtc tttggactct
     2161 caggaatctt tcagatgctg caactaaaca ggaagggatg gaaggtctcc ttgggactct
     2221 tgttcagctt ctgggttcag atgatataaa tgtggtcacc tgtgcagctg gaattctttc
     2281 taacctcact tgcaataatt ataagaacaa gatgatggtc tgccaagtgg gtggtataga
     2341 ggctcttgtg cgtactgtcc ttcgggctgg tgacagggaa gacatcactg agcctgccat
     2401 ctgtgctctt cgtcatctga ccagccgaca ccaagaagca gagatggccc agaatgcagt
     2461 tcgccttcac tatggactac cagttgtggt taagctctta cacccaccat cccactggcc
     2521 tctgataaag gctactgttg gattgattcg aaatcttgcc ctttgtcccg caaatcatgc
     2581 acctttgcgt gagcagggtg ccattccacg actagttcag ttgcttgttc gtgcacatca
     2641 ggatacccag cgccgtacgt ccatgggtgg gacacagcag caatttgtgg agggggtccg
     2701 catggaagaa atagttgaag gttgtaccgg agcccttcac atcctagctc gggatgttca
     2761 caaccgaatt gttatcagag gactaaatac cattccattg tttgtgcagc tgctttattc
     2821 tcccattgaa aacatccaaa gagtagctgc aggggtcctc tgtgaacttg ctcaggacaa
     2881 ggaagctgca gaagctattg aagctgaggg agccacagct cctctgacag agttacttca
     2941 ctctaggaat gaaggtgtgg cgacatatgc agctgctgtt ttgttccgaa tgtctgagga
     3001 caagccacaa gattacaaga aacggctttc agttgagctg accagctctc tcttcagaac
     3061 agagccaatg gcttggaatg agactgctga tcttggactt gatattggtg cccagggaga
     3121 accccttgga tatcgccagg atgatcctag ctatcgttct tttcactctg gtggatatgg
     3181 ccaggatgcc ttgggtatgg accccatgat ggaacatgag atgggtggcc accaccctgg
     3241 tgctgactat ccagttgatg ggctgccaga tctggggcat gcccaggacc tcatggatgg
     3301 gctgcctcca ggtgacagca atcagctggc ctggtttgat actgacctgt aatctagagt
     3361 cgacccgggc ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat
     3421 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3481 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3541 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     3601 gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa
     3661 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3721 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3841 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4081 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4141 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     4201 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     4261 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     4681 gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc
     4741 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4801 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4861 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4921 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4981 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5041 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5101 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5161 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5221 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5281 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5341 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5401 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5461 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5521 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     5581 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc
     5641 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5701 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5761 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5821 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5881 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5941 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6001 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6061 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6121 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6181 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6241 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6301 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6361 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6421 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6481 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6541 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6601 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6661 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6721 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6781 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6841 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6901 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6961 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7681 tggccttttg ctcacatggc tcgacagatc gatcttcaat attggccatt agccatatta
     7741 ttcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     7801 tatcataata t
//
