LOCUS Exported 5139 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway Donor vector containing CTNNB1,part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_CTNNB1_p.R535Q 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 13, 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" rep_origin 21..609 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 510..529 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 763..780 /label=L4440 /note="L4440 vector, forward primer" terminator 939..966 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 1058..1144 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 1193..1215 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1207..1224 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1208..1224 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1240..1339 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 1342..3684 /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 ALCPANHAPLREQGAIPQLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL PDLGHAQDLMDGLPPGDSNQLAWFDTDL" protein_bind complement(3689..3788) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3805..3824) /label=T7 /note="T7 promoter, forward primer" promoter complement(3806..3824) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3829..3845) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3829..3845) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4276..5067 /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" ORIGIN 1 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 61 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 781 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 841 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 901 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg 961 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg 1021 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt 1081 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta 1141 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca 1201 cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga 1261 ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact 1321 ttgtacaaaa aagttggcac catggctact caagctgatt tgatggagtt ggacatggcc 1381 atggaaccag acagaaaagc ggctgttagt cactggcagc aacagtctta cctggactct 1441 ggaatccatt ctggtgccac taccacagct ccttctctga gtggtaaagg caatcctgag 1501 gaagaggatg tggatacctc ccaagtcctg tatgagtggg aacagggatt ttctcagtcc 1561 ttcactcaag aacaagtagc tgatattgat ggacagtatg caatgactcg agctcagagg 1621 gtacgagctg ctatgttccc tgagacatta gatgagggca tgcagatccc atctacacag 1681 tttgatgctg ctcatcccac taatgtccag cgtttggctg aaccatcaca gatgctgaaa 1741 catgcagttg taaacttgat taactatcaa gatgatgcag aacttgccac acgtgcaatc 1801 cctgaactga caaaactgct aaatgacgag gaccaggtgg tggttaataa ggctgcagtt 1861 atggtccatc agctttctaa aaaggaagct tccagacacg ctatcatgcg ttctcctcag 1921 atggtgtctg ctattgtacg taccatgcag aatacaaatg atgtagaaac agctcgttgt 1981 accgctggga ccttgcataa cctttcccat catcgtgagg gcttactggc catctttaag 2041 tctggaggca ttcctgccct ggtgaaaatg cttggttcac cagtggattc tgtgttgttt 2101 tatgccatta caactctcca caacctttta ttacatcaag aaggagctaa aatggcagtg 2161 cgtttagctg gtgggctgca gaaaatggtt gccttgctca acaaaacaaa tgttaaattc 2221 ttggctatta cgacagactg ccttcaaatt ttagcttatg gcaaccaaga aagcaagctc 2281 atcatactgg ctagtggtgg accccaagct ttagtaaata taatgaggac ctatacttac 2341 gaaaaactac tgtggaccac aagcagagtg ctgaaggtgc tatctgtctg ctctagtaat 2401 aagccggcta ttgtagaagc tggtggaatg caagctttag gacttcacct gacagatcca 2461 agtcaacgtc ttgttcagaa ctgtctttgg actctcagga atctttcaga tgctgcaact 2521 aaacaggaag ggatggaagg tctccttggg actcttgttc agcttctggg ttcagatgat 2581 ataaatgtgg tcacctgtgc agctggaatt ctttctaacc tcacttgcaa taattataag 2641 aacaagatga tggtctgcca agtgggtggt atagaggctc ttgtgcgtac tgtccttcgg 2701 gctggtgaca gggaagacat cactgagcct gccatctgtg ctcttcgtca tctgaccagc 2761 cgacaccaag aagcagagat ggcccagaat gcagttcgcc ttcactatgg actaccagtt 2821 gtggttaagc tcttacaccc accatcccac tggcctctga taaaggctac tgttggattg 2881 attcgaaatc ttgccctttg tcccgcaaat catgcacctt tgcgtgagca gggtgccatt 2941 ccacaactag ttcagttgct tgttcgtgca catcaggata cccagcgccg tacgtccatg 3001 ggtgggacac agcagcaatt tgtggagggg gtccgcatgg aagaaatagt tgaaggttgt 3061 accggagccc ttcacatcct agctcgggat gttcacaacc gaattgttat cagaggacta 3121 aataccattc cattgtttgt gcagctgctt tattctccca ttgaaaacat ccaaagagta 3181 gctgcagggg tcctctgtga acttgctcag gacaaggaag ctgcagaagc tattgaagct 3241 gagggagcca cagctcctct gacagagtta cttcactcta ggaatgaagg tgtggcgaca 3301 tatgcagctg ctgttttgtt ccgaatgtct gaggacaagc cacaagatta caagaaacgg 3361 ctttcagttg agctgaccag ctctctcttc agaacagagc caatggcttg gaatgagact 3421 gctgatcttg gacttgatat tggtgcccag ggagaacccc ttggatatcg ccaggatgat 3481 cctagctatc gttcttttca ctctggtgga tatggccagg atgccttggg tatggacccc 3541 atgatggaac atgagatggg tggccaccac cctggtgctg actatccagt tgatgggctg 3601 ccagatctgg ggcatgccca ggacctcatg gatgggctgc ctccaggtga cagcaatcag 3661 ctggcctggt ttgatactga cctgtgattg ccaactttct tgtacaaagt tggcattata 3721 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc 3781 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt 3841 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat 3901 atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc 3961 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa 4021 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga 4081 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac 4141 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg 4201 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa 4261 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag 4321 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc 4381 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc 4441 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct 4501 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac 4561 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat 4621 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg 4681 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat 4741 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg 4801 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac 4861 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc 4921 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa 4981 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc 5041 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac 5101 gtgagttacg cgtcgttcca ctgagcgtca gaccccgta //