LOCUS       sequence-168801-        5139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing , part of the Target Accelerator 
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_CTNNB1_p.D162E
FEATURES             Location/Qualifiers
     source          1..5139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             671..3013
                     /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
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNEE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     protein_bind    complement(3018..3117)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3134..3153)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3135..3153)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3158..3174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3158..3174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3605..4396
                     /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      4489..5077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4978..4997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atggctactc aagctgattt gatggagttg gacatggcca tggaaccaga
      721 cagaaaagcg gctgttagtc actggcagca acagtcttac ctggactctg gaatccattc
      781 tggtgccact accacagctc cttctctgag tggtaaaggc aatcctgagg aagaggatgt
      841 ggatacctcc caagtcctgt atgagtggga acagggattt tctcagtcct tcactcaaga
      901 acaagtagct gatattgatg gacagtatgc aatgactcga gctcagaggg tacgagctgc
      961 tatgttccct gagacattag atgagggcat gcagatccca tctacacagt ttgatgctgc
     1021 tcatcccact aatgtccagc gtttggctga accatcacag atgctgaaac atgcagttgt
     1081 aaacttgatt aactatcaag atgatgcaga acttgccaca cgtgcaatcc ctgaactgac
     1141 aaaactgcta aatgaagagg accaggtggt ggttaataag gctgcagtta tggtccatca
     1201 gctttctaaa aaggaagctt ccagacacgc tatcatgcgt tctcctcaga tggtgtctgc
     1261 tattgtacgt accatgcaga atacaaatga tgtagaaaca gctcgttgta ccgctgggac
     1321 cttgcataac ctttcccatc atcgtgaggg cttactggcc atctttaagt ctggaggcat
     1381 tcctgccctg gtgaaaatgc ttggttcacc agtggattct gtgttgtttt atgccattac
     1441 aactctccac aaccttttat tacatcaaga aggagctaaa atggcagtgc gtttagctgg
     1501 tgggctgcag aaaatggttg ccttgctcaa caaaacaaat gttaaattct tggctattac
     1561 gacagactgc cttcaaattt tagcttatgg caaccaagaa agcaagctca tcatactggc
     1621 tagtggtgga ccccaagctt tagtaaatat aatgaggacc tatacttacg aaaaactact
     1681 gtggaccaca agcagagtgc tgaaggtgct atctgtctgc tctagtaata agccggctat
     1741 tgtagaagct ggtggaatgc aagctttagg acttcacctg acagatccaa gtcaacgtct
     1801 tgttcagaac tgtctttgga ctctcaggaa tctttcagat gctgcaacta aacaggaagg
     1861 gatggaaggt ctccttggga ctcttgttca gcttctgggt tcagatgata taaatgtggt
     1921 cacctgtgca gctggaattc tttctaacct cacttgcaat aattataaga acaagatgat
     1981 ggtctgccaa gtgggtggta tagaggctct tgtgcgtact gtccttcggg ctggtgacag
     2041 ggaagacatc actgagcctg ccatctgtgc tcttcgtcat ctgaccagcc gacaccaaga
     2101 agcagagatg gcccagaatg cagttcgcct tcactatgga ctaccagttg tggttaagct
     2161 cttacaccca ccatcccact ggcctctgat aaaggctact gttggattga ttcgaaatct
     2221 tgccctttgt cccgcaaatc atgcaccttt gcgtgagcag ggtgccattc cacgactagt
     2281 tcagttgctt gttcgtgcac atcaggatac ccagcgccgt acgtccatgg gtgggacaca
     2341 gcagcaattt gtggaggggg tccgcatgga agaaatagtt gaaggttgta ccggagccct
     2401 tcacatccta gctcgggatg ttcacaaccg aattgttatc agaggactaa ataccattcc
     2461 attgtttgtg cagctgcttt attctcccat tgaaaacatc caaagagtag ctgcaggggt
     2521 cctctgtgaa cttgctcagg acaaggaagc tgcagaagct attgaagctg agggagccac
     2581 agctcctctg acagagttac ttcactctag gaatgaaggt gtggcgacat atgcagctgc
     2641 tgttttgttc cgaatgtctg aggacaagcc acaagattac aagaaacggc tttcagttga
     2701 gctgaccagc tctctcttca gaacagagcc aatggcttgg aatgagactg ctgatcttgg
     2761 acttgatatt ggtgcccagg gagaacccct tggatatcgc caggatgatc ctagctatcg
     2821 ttcttttcac tctggtggat atggccagga tgccttgggt atggacccca tgatggaaca
     2881 tgagatgggt ggccaccacc ctggtgctga ctatccagtt gatgggctgc cagatctggg
     2941 gcatgcccag gacctcatgg atgggctgcc tccaggtgac agcaatcagc tggcctggtt
     3001 tgatactgac ctgtgattgc caactttctt gtacaaagtt ggcattataa gaaagcattg
     3061 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3121 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     3181 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     3241 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     3301 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     3361 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     3421 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     3481 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     3541 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     3601 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     3661 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     3721 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     3781 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     3841 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     3901 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     3961 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     4021 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     4081 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     4141 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     4201 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     4261 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     4321 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     4381 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     4441 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4501 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4561 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4621 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4681 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4741 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4801 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4861 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4921 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4981 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5041 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5101 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
