LOCUS       sequence-167015-        4561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PPP2R1A , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4561)
  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 4561)
  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_PPP2R1A_p.R258H
FEATURES             Location/Qualifiers
     source          1..4561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      190..199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1961..2060)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2077..2096)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2078..2096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2101..2117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2101..2117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2548..3339
                     /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      3432..4020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3921..3940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4174..4191
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4350..4377)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4469..4555)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catggcggcg gccgacggcg acgactcgct gtaccccatc gcggtgctca
      241 tagacgaact ccgcaatgag gacgttcagc ttcgcctcaa cagcatcaag aagctgtcca
      301 ccatcgcctt ggcccttggg gttgaaagga cccgaagtga gcttctgcct ttccttacag
      361 ataccatcta tgatgaagat gaggtcctcc tggccctggc agaacagctg ggaaccttca
      421 ctaccctggt gggaggccca gagtacgtgc actgcctgct gccaccgctg gagtcgctgg
      481 ccacagtgga ggagacagtg gtgcgggaca aggcagtgga gtccttacgg gccatctcac
      541 acgagcactc gccctctgac ctggaggcgc actttgtgcc gctagtgaag cggctggcgg
      601 gcggcgactg gttcacctcc cgcacctcgg cctgcggcct cttctccgtc tgctaccccc
      661 gagtgtccag tgctgtgaag gcggaacttc gacagtactt ccggaacctg tgctcagatg
      721 acacccccat ggtgcggcgg gccgcagcct ccaagctggg ggagtttgcc aaggtgctgg
      781 agctggacaa cgtcaagagt gagatcatcc ccatgttctc caacctggcc tctgacgagc
      841 aggactcggt gcggctgctg gcggtggagg cgtgcgtgaa catcgcccag cttctgcccc
      901 aggaggatct ggaggccctg gtgatgccca ctctgcgcca ggccgctgaa gacaagtcct
      961 ggcacgtccg ctacatggtg gctgacaagt tcacagagct ccagaaagca gtggggcctg
     1021 agatcaccaa gacagacctg gtccctgcct tccagaacct gatgaaagac tgtgaggccg
     1081 aggtgagggc cgcagcctcc cacaaggtca aagagttctg tgaaaacctc tcagctgact
     1141 gtcgggagaa tgtgatcatg tcccagatct tgccctgcat caaggagctg gtgtccgatg
     1201 ccaaccaaca tgtcaagtct gccctggcct cagtcatcat gggtctctct cccatcttgg
     1261 gcaaagacaa caccatcgag cacctcttgc ccctcttcct ggctcagctg aaggatgagt
     1321 gccctgaggt acggctgaac atcatctcta acctggactg tgtgaacgag gtgattggca
     1381 tccggcagct gtcccagtcc ctgctccctg ccattgtgga gctggctgag gacgccaagt
     1441 ggcgggtgcg gctggccatc attgagtaca tgcccctcct ggctggacag ctgggagtgg
     1501 agttctttga tgagaaactt aactccttgt gcatggcctg gcttgtggat catgtatatg
     1561 ccatccgcga ggcagccacc agcaacctga agaagctagt ggaaaagttt gggaaggagt
     1621 gggcccatgc cacaatcatc cccaaggtct tggccatgtc cggagacccc aactacctgc
     1681 accgcatgac tacgctcttc tgcatcaatg tgctgtctga ggtctgtggg caggacatca
     1741 ccaccaagca catgctaccc acggttctgc gcatggctgg ggacccggtt gccaatgtcc
     1801 gcttcaatgt ggccaagtct ctgcagaaga tagggcccat cctggacaac agcaccttgc
     1861 agagtgaagt caagcccatc ctagagaagc tgacccagga ccaggatgtg gacgtcaaat
     1921 actttgccca ggaggctctg actgttctgt ctcgcctgat tgccaacttt cttgtacaaa
     1981 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     2041 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     2101 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2161 caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact
     2221 tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc
     2281 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
     2341 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     2401 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
     2461 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
     2521 agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact
     2581 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
     2641 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
     2701 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     2761 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     2821 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     2881 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     2941 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     3001 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     3061 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     3121 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     3181 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     3241 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     3301 tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca
     3361 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4201 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4261 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     4321 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     4381 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     4441 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     4501 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     4561 t
//
