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_p.T257I
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"
     CDS             329..1120
                     /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      1213..1801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1702..1721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1955..1972
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2131..2158
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2250..2336
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2385..2407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2399..2416
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2400..2416
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2432..2531
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2534..4879
                     /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
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAIITLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     protein_bind    complement(4881..4980)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4997..5016)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4998..5016)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5021..5037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5021..5037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
       61 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
      121 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
      181 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
      241 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
      301 cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac
      361 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
      421 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
      481 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
      541 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
      601 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
      661 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
      721 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
      781 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
      841 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
      901 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
      961 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     1021 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     1081 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     1141 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     1201 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1261 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1321 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1381 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1441 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1501 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1561 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1621 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1681 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1741 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1801 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1861 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1921 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1981 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2041 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     2101 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     2161 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     2221 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     2281 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     2341 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
     2401 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
     2461 gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa
     2521 aaaagttggc accatggcta ctcaagctga tttgatggag ttggacatgg ccatggaacc
     2581 agacagaaaa gcggctgtta gtcactggca gcaacagtct tacctggact ctggaatcca
     2641 ttctggtgcc actaccacag ctccttctct gagtggtaaa ggcaatcctg aggaagagga
     2701 tgtggatacc tcccaagtcc tgtatgagtg ggaacaggga ttttctcagt ccttcactca
     2761 agaacaagta gctgatattg atggacagta tgcaatgact cgagctcaga gggtacgagc
     2821 tgctatgttc cctgagacat tagatgaggg catgcagatc ccatctacac agtttgatgc
     2881 tgctcatccc actaatgtcc agcgtttggc tgaaccatca cagatgctga aacatgcagt
     2941 tgtaaacttg attaactatc aagatgatgc agaacttgcc acacgtgcaa tccctgaact
     3001 gacaaaactg ctaaatgacg aggaccaggt ggtggttaat aaggctgcag ttatggtcca
     3061 tcagctttct aaaaaggaag cttccagaca cgctatcatg cgttctcctc agatggtgtc
     3121 tgctattgta cgtaccatgc agaatacaaa tgatgtagaa acagctcgtt gtaccgctgg
     3181 gaccttgcat aacctttccc atcatcgtga gggcttactg gccatcttta agtctggagg
     3241 cattcctgcc ctggtgaaaa tgcttggttc accagtggat tctgtgttgt tttatgccat
     3301 tataactctc cacaaccttt tattacatca agaaggagct aaaatggcag tgcgtttagc
     3361 tggtgggctg cagaaaatgg ttgccttgct caacaaaaca aatgttaaat tcttggctat
     3421 tacgacagac tgccttcaaa ttttagctta tggcaaccaa gaaagcaagc tcatcatact
     3481 ggctagtggt ggaccccaag ctttagtaaa tataatgagg acctatactt acgaaaaact
     3541 actgtggacc acaagcagag tgctgaaggt gctatctgtc tgctctagta ataagccggc
     3601 tattgtagaa gctggtggaa tgcaagcttt aggacttcac ctgacagatc caagtcaacg
     3661 tcttgttcag aactgtcttt ggactctcag gaatctttca gatgctgcaa ctaaacagga
     3721 agggatggaa ggtctccttg ggactcttgt tcagcttctg ggttcagatg atataaatgt
     3781 ggtcacctgt gcagctggaa ttctttctaa cctcacttgc aataattata agaacaagat
     3841 gatggtctgc caagtgggtg gtatagaggc tcttgtgcgt actgtccttc gggctggtga
     3901 cagggaagac atcactgagc ctgccatctg tgctcttcgt catctgacca gccgacacca
     3961 agaagcagag atggcccaga atgcagttcg ccttcactat ggactaccag ttgtggttaa
     4021 gctcttacac ccaccatccc actggcctct gataaaggct actgttggat tgattcgaaa
     4081 tcttgccctt tgtcccgcaa atcatgcacc tttgcgtgag cagggtgcca ttccacgact
     4141 agttcagttg cttgttcgtg cacatcagga tacccagcgc cgtacgtcca tgggtgggac
     4201 acagcagcaa tttgtggagg gggtccgcat ggaagaaata gttgaaggtt gtaccggagc
     4261 ccttcacatc ctagctcggg atgttcacaa ccgaattgtt atcagaggac taaataccat
     4321 tccattgttt gtgcagctgc tttattctcc cattgaaaac atccaaagag tagctgcagg
     4381 ggtcctctgt gaacttgctc aggacaagga agctgcagaa gctattgaag ctgagggagc
     4441 cacagctcct ctgacagagt tacttcactc taggaatgaa ggtgtggcga catatgcagc
     4501 tgctgttttg ttccgaatgt ctgaggacaa gccacaagat tacaagaaac ggctttcagt
     4561 tgagctgacc agctctctct tcagaacaga gccaatggct tggaatgaga ctgctgatct
     4621 tggacttgat attggtgccc agggagaacc ccttggatat cgccaggatg atcctagcta
     4681 tcgttctttt cactctggtg gatatggcca ggatgccttg ggtatggacc ccatgatgga
     4741 acatgagatg ggtggccacc accctggtgc tgactatcca gttgatgggc tgccagatct
     4801 ggggcatgcc caggacctca tggatgggct gcctccaggt gacagcaatc agctggcctg
     4861 gtttgatact gacctgtaat tgccaacttt cttgtacaaa gttggcatta taagaaagca
     4921 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     4981 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     5041 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     5101 atgcctcctc tagaccagcc aggacagaaa tgcctcgac
//
