LOCUS Exported 5139 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing CTNNB1 , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_CTNNB1_WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5139) AUTHORS Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn WC TITLE Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles. JOURNAL Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4. PUBMED 27147599 REFERENCE 2 (bases 1 to 5139) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5139 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 174..201 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 293..379 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 428..450 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 442..459 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 443..459 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 475..574 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 577..2922 /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" CDS 577..2919 /codon_start=1 /gene="Homo sapiens CTNNB1" /product="human cadherin-associated protein that also plays a role in the canonical Wnt signaling pathway" /label=-beta-catenin /note="also known as catenin beta-1" /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL PDLGHAQDLMDGLPPGDSNQLAWFDTDL" protein_bind complement(2924..3023) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3040..3059) /label=T7 /note="T7 promoter, forward primer" promoter complement(3041..3059) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3064..3080) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3064..3080) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 3511..4302 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 4395..4983 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4884..4903 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5137..15 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt 61 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 121 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc 181 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg 241 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc 301 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact 361 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg 421 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccccaaata 481 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgatga gcaatgcttt 541 tttataatgc caactttgta caaaaaagtt ggcaccatgg ctactcaagc tgatttgatg 601 gagttggaca tggccatgga accagacaga aaagcggctg ttagtcactg gcagcaacag 661 tcttacctgg actctggaat ccattctggt gccactacca cagctccttc tctgagtggt 721 aaaggcaatc ctgaggaaga ggatgtggat acctcccaag tcctgtatga gtgggaacag 781 ggattttctc agtccttcac tcaagaacaa gtagctgata ttgatggaca gtatgcaatg 841 actcgagctc agagggtacg agctgctatg ttccctgaga cattagatga gggcatgcag 901 atcccatcta cacagtttga tgctgctcat cccactaatg tccagcgttt ggctgaacca 961 tcacagatgc tgaaacatgc agttgtaaac ttgattaact atcaagatga tgcagaactt 1021 gccacacgtg caatccctga actgacaaaa ctgctaaatg acgaggacca ggtggtggtt 1081 aataaggctg cagttatggt ccatcagctt tctaaaaagg aagcttccag acacgctatc 1141 atgcgttctc ctcagatggt gtctgctatt gtacgtacca tgcagaatac aaatgatgta 1201 gaaacagctc gttgtaccgc tgggaccttg cataaccttt cccatcatcg tgagggctta 1261 ctggccatct ttaagtctgg aggcattcct gccctggtga aaatgcttgg ttcaccagtg 1321 gattctgtgt tgttttatgc cattacaact ctccacaacc ttttattaca tcaagaagga 1381 gctaaaatgg cagtgcgttt agctggtggg ctgcagaaaa tggttgcctt gctcaacaaa 1441 acaaatgtta aattcttggc tattacgaca gactgccttc aaattttagc ttatggcaac 1501 caagaaagca agctcatcat actggctagt ggtggacccc aagctttagt aaatataatg 1561 aggacctata cttacgaaaa actactgtgg accacaagca gagtgctgaa ggtgctatct 1621 gtctgctcta gtaataagcc ggctattgta gaagctggtg gaatgcaagc tttaggactt 1681 cacctgacag atccaagtca acgtcttgtt cagaactgtc tttggactct caggaatctt 1741 tcagatgctg caactaaaca ggaagggatg gaaggtctcc ttgggactct tgttcagctt 1801 ctgggttcag atgatataaa tgtggtcacc tgtgcagctg gaattctttc taacctcact 1861 tgcaataatt ataagaacaa gatgatggtc tgccaagtgg gtggtataga ggctcttgtg 1921 cgtactgtcc ttcgggctgg tgacagggaa gacatcactg agcctgccat ctgtgctctt 1981 cgtcatctga ccagccgaca ccaagaagca gagatggccc agaatgcagt tcgccttcac 2041 tatggactac cagttgtggt taagctctta cacccaccat cccactggcc tctgataaag 2101 gctactgttg gattgattcg aaatcttgcc ctttgtcccg caaatcatgc acctttgcgt 2161 gagcagggtg ccattccacg actagttcag ttgcttgttc gtgcacatca ggatacccag 2221 cgccgtacgt ccatgggtgg gacacagcag caatttgtgg agggggtccg catggaagaa 2281 atagttgaag gttgtaccgg agcccttcac atcctagctc gggatgttca caaccgaatt 2341 gttatcagag gactaaatac cattccattg tttgtgcagc tgctttattc tcccattgaa 2401 aacatccaaa gagtagctgc aggggtcctc tgtgaacttg ctcaggacaa ggaagctgca 2461 gaagctattg aagctgaggg agccacagct cctctgacag agttacttca ctctaggaat 2521 gaaggtgtgg cgacatatgc agctgctgtt ttgttccgaa tgtctgagga caagccacaa 2581 gattacaaga aacggctttc agttgagctg accagctctc tcttcagaac agagccaatg 2641 gcttggaatg agactgctga tcttggactt gatattggtg cccagggaga accccttgga 2701 tatcgccagg atgatcctag ctatcgttct tttcactctg gtggatatgg ccaggatgcc 2761 ttgggtatgg accccatgat ggaacatgag atgggtggcc accaccctgg tgctgactat 2821 ccagttgatg ggctgccaga tctggggcat gcccaggacc tcatggatgg gctgcctcca 2881 ggtgacagca atcagctggc ctggtttgat actgacctgt aattgccaac tttcttgtac 2941 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc 3001 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac 3061 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt 3121 gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg 3181 acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga 3241 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca 3301 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc 3361 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa 3421 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag 3481 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg 3541 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc 3601 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg 3661 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt 3721 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt 3781 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga 3841 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat 3901 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg 3961 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta 4021 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg 4081 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac 4141 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct 4201 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc 4261 cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga 4321 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag 4381 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 4441 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 4501 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 4561 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 4621 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 4681 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 4741 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 4801 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 4861 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 4921 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 4981 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 5041 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 5101 gctgataccg ctcgccgcag ccgaacgacc gagcgcagc //