LOCUS       sequence-168630-        6000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PIK3CA, part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  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 6000)
  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_PIK3CA_p.D1045V
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      22..610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     511..530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     764..781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(940..967)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1059..1145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1194..1216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1208..1225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1209..1225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1241..1340
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4551..4650)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4667..4686)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4668..4686)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4691..4707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4691..4707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5138..5929
                     /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 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
       61 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      721 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      781 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      841 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      901 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
      961 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     1021 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     1081 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     1141 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc
     1201 acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg
     1261 actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac
     1321 tttgtacaaa aaagttggca ccatgcctcc acgaccatca tcaggtgaac tgtggggcat
     1381 ccacttgatg cccccaagaa tcctagtaga atgtttacta ccaaatggaa tgatagtgac
     1441 tttagaatgc ctccgtgagg ctacattaat aaccataaag catgaactat ttaaagaagc
     1501 aagaaaatac cccctccatc aacttcttca agatgaatct tcttacattt tcgtaagtgt
     1561 tactcaagaa gcagaaaggg aagaattttt tgatgaaaca agacgacttt gtgaccttcg
     1621 gctttttcaa ccctttttaa aagtaattga accagtaggc aaccgtgaag aaaagatcct
     1681 caatcgagaa attggttttg ctatcggcat gccagtgtgt gaatttgata tggttaaaga
     1741 tccagaagta caggacttcc gaagaaatat tctgaacgtt tgtaaagaag ctgtggatct
     1801 tagggacctc aattcacctc atagtagagc aatgtatgtc tatcctccaa atgtagaatc
     1861 ttcaccagaa ttgccaaagc acatatataa taaattagat aaagggcaaa taatagtggt
     1921 gatctgggta atagtttctc caaataatga caagcagaag tatactctga aaatcaacca
     1981 tgactgtgta ccagaacaag taattgctga agcaatcagg aaaaaaactc gaagtatgtt
     2041 gctatcctct gaacaactaa aactctgtgt tttagaatat cagggcaagt atattttaaa
     2101 agtgtgtgga tgtgatgaat acttcctaga aaaatatcct ctgagtcagt ataagtatat
     2161 aagaagctgt ataatgcttg ggaggatgcc caatttgatg ttgatggcta aagaaagcct
     2221 ttattctcaa ctgccaatgg actgttttac aatgccatct tattccagac gcatttccac
     2281 agctacacca tatatgaatg gagaaacatc tacaaaatcc ctttgggtta taaatagtgc
     2341 actcagaata aaaattcttt gtgcaaccta cgtgaatgta aatattcgag acattgataa
     2401 gatctatgtt cgaacaggta tctaccatgg aggagaaccc ttatgtgaca atgtgaacac
     2461 tcaaagagta ccttgttcca atcccaggtg gaatgaatgg ctgaattatg atatatacat
     2521 tcctgatctt cctcgtgctg ctcgactttg cctttccatt tgctctgtta aaggccgaaa
     2581 gggtgctaaa gaggaacact gtccattggc atggggaaat ataaacttgt ttgattacac
     2641 agacactcta gtatctggaa aaatggcttt gaatctttgg ccagtacctc atggattaga
     2701 agatttgctg aaccctattg gtgttactgg atcaaatcca aataaagaaa ctccatgctt
     2761 agagttggag tttgactggt tcagcagtgt ggtaaagttc ccagatatgt cagtgattga
     2821 agagcatgcc aattggtctg tatcccgaga agcaggattt agctattccc acgcaggact
     2881 gagtaacaga ctagctagag acaatgaatt aagggaaaat gacaaagaac agctcaaagc
     2941 aatttctaca cgagatcctc tctctgaaat cactgagcag gagaaagatt ttctatggag
     3001 tcacagacac tattgtgtaa ctatccccga aattctaccc aaattgcttc tgtctgttaa
     3061 atggaattct agagatgaag tagcccagat gtattgcttg gtaaaagatt ggcctccaat
     3121 caaacctgaa caggctatgg aacttctgga ctgtaattac ccagatccta tggttcgagg
     3181 ttttgctgtt cggtgcttgg aaaaatattt aacagatgac aaactttctc agtatttaat
     3241 tcagctagta caggtcctaa aatatgaaca atatttggat aacttgcttg tgagattttt
     3301 actgaagaaa gcattgacta atcaaaggat tgggcacttt ttcttttggc atttaaaatc
     3361 tgagatgcac aataaaacag ttagccagag gtttggcctg cttttggagt cctattgtcg
     3421 tgcatgtggg atgtatttga agcacctgaa taggcaagtc gaggcaatgg aaaagctcat
     3481 taacttaact gacattctca aacaggagaa gaaggatgaa acacaaaagg tacagatgaa
     3541 gtttttagtt gagcaaatga ggcgaccaga tttcatggat gctctacagg gctttctgtc
     3601 tcctctaaac cctgctcatc aactaggaaa cctcaggctt gaagagtgtc gaattatgtc
     3661 ctctgcaaaa aggccactgt ggttgaattg ggagaaccca gacatcatgt cagagttact
     3721 gtttcagaac aatgagatca tctttaaaaa tggggatgat ttacggcaag atatgctaac
     3781 acttcaaatt attcgtatta tggaaaatat ctggcaaaat caaggtcttg atcttcgaat
     3841 gttaccttat ggttgtctgt caatcggtga ctgtgtggga cttattgagg tggtgcgaaa
     3901 ttctcacact attatgcaaa ttcagtgcaa aggcggcttg aaaggtgcac tgcagttcaa
     3961 cagccacaca ctacatcagt ggctcaaaga caagaacaaa ggagaaatat atgatgcagc
     4021 cattgacctg tttacacgtt catgtgctgg atactgtgta gctaccttca ttttgggaat
     4081 tggagatcgt cacaatagta acatcatggt gaaagacgat ggacaactgt ttcatataga
     4141 ttttggacac tttttggatc acaagaagaa aaaatttggt tataaacgag aacgtgtgcc
     4201 atttgttttg acacaggatt tcttaatagt gattagtaaa ggagcccaag aatgcacaaa
     4261 gacaagagaa tttgagaggt ttcaggagat gtgttacaag gcttatctag ctattcgaca
     4321 gcatgccaat ctcttcataa atcttttctc aatgatgctt ggctctggaa tgccagaact
     4381 acaatctttt gatgacattg catacattcg aaagacccta gccttagata aaactgagca
     4441 agaggctttg gagtatttca tgaaacaaat gaatgttgca catcatggtg gctggacaac
     4501 aaaaatggat tggatcttcc acacaattaa acagcatgca ttgaactgat tgccaacttt
     4561 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     4621 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     4681 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     4741 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa
     4801 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     4861 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     4921 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     4981 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     5041 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     5101 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     5161 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     5221 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     5281 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     5341 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     5401 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     5461 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     5521 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     5581 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     5641 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     5701 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     5761 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     5821 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     5881 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     5941 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
//
