LOCUS       sequence-162438-        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   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
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: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_PIK3CA_p.E545K
FEATURES             Location/Qualifiers
     source          1..6000
                     /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"
     protein_bind    complement(3400..3499)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3516..3535)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3517..3535)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3540..3556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3540..3556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3987..4778
                     /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      4871..5459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5360..5379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5613..5630
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5789..5816)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5908..5994)
                     /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 catgcctcca cgaccatcat caggtgaact gtggggcatc cacttgatgc
      241 ccccaagaat cctagtagaa tgtttactac caaatggaat gatagtgact ttagaatgcc
      301 tccgtgaggc tacattaata accataaagc atgaactatt taaagaagca agaaaatacc
      361 ccctccatca acttcttcaa gatgaatctt cttacatttt cgtaagtgtt actcaagaag
      421 cagaaaggga agaatttttt gatgaaacaa gacgactttg tgaccttcgg ctttttcaac
      481 cctttttaaa agtaattgaa ccagtaggca accgtgaaga aaagatcctc aatcgagaaa
      541 ttggttttgc tatcggcatg ccagtgtgtg aatttgatat ggttaaagat ccagaagtac
      601 aggacttccg aagaaatatt ctgaacgttt gtaaagaagc tgtggatctt agggacctca
      661 attcacctca tagtagagca atgtatgtct atcctccaaa tgtagaatct tcaccagaat
      721 tgccaaagca catatataat aaattagata aagggcaaat aatagtggtg atctgggtaa
      781 tagtttctcc aaataatgac aagcagaagt atactctgaa aatcaaccat gactgtgtac
      841 cagaacaagt aattgctgaa gcaatcagga aaaaaactcg aagtatgttg ctatcctctg
      901 aacaactaaa actctgtgtt ttagaatatc agggcaagta tattttaaaa gtgtgtggat
      961 gtgatgaata cttcctagaa aaatatcctc tgagtcagta taagtatata agaagctgta
     1021 taatgcttgg gaggatgccc aatttgatgt tgatggctaa agaaagcctt tattctcaac
     1081 tgccaatgga ctgttttaca atgccatctt attccagacg catttccaca gctacaccat
     1141 atatgaatgg agaaacatct acaaaatccc tttgggttat aaatagtgca ctcagaataa
     1201 aaattctttg tgcaacctac gtgaatgtaa atattcgaga cattgataag atctatgttc
     1261 gaacaggtat ctaccatgga ggagaaccct tatgtgacaa tgtgaacact caaagagtac
     1321 cttgttccaa tcccaggtgg aatgaatggc tgaattatga tatatacatt cctgatcttc
     1381 ctcgtgctgc tcgactttgc ctttccattt gctctgttaa aggccgaaag ggtgctaaag
     1441 aggaacactg tccattggca tggggaaata taaacttgtt tgattacaca gacactctag
     1501 tatctggaaa aatggctttg aatctttggc cagtacctca tggattagaa gatttgctga
     1561 accctattgg tgttactgga tcaaatccaa ataaagaaac tccatgctta gagttggagt
     1621 ttgactggtt cagcagtgtg gtaaagttcc cagatatgtc agtgattgaa gagcatgcca
     1681 attggtctgt atcccgagaa gcaggattta gctattccca cgcaggactg agtaacagac
     1741 tagctagaga caatgaatta agggaaaatg acaaagaaca gctcaaagca atttctacac
     1801 gagatcctct ctctgaaatc actaagcagg agaaagattt tctatggagt cacagacact
     1861 attgtgtaac tatccccgaa attctaccca aattgcttct gtctgttaaa tggaattcta
     1921 gagatgaagt agcccagatg tattgcttgg taaaagattg gcctccaatc aaacctgaac
     1981 aggctatgga acttctggac tgtaattacc cagatcctat ggttcgaggt tttgctgttc
     2041 ggtgcttgga aaaatattta acagatgaca aactttctca gtatttaatt cagctagtac
     2101 aggtcctaaa atatgaacaa tatttggata acttgcttgt gagattttta ctgaagaaag
     2161 cattgactaa tcaaaggatt gggcactttt tcttttggca tttaaaatct gagatgcaca
     2221 ataaaacagt tagccagagg tttggcctgc ttttggagtc ctattgtcgt gcatgtggga
     2281 tgtatttgaa gcacctgaat aggcaagtcg aggcaatgga aaagctcatt aacttaactg
     2341 acattctcaa acaggagaag aaggatgaaa cacaaaaggt acagatgaag tttttagttg
     2401 agcaaatgag gcgaccagat ttcatggatg ctctacaggg ctttctgtct cctctaaacc
     2461 ctgctcatca actaggaaac ctcaggcttg aagagtgtcg aattatgtcc tctgcaaaaa
     2521 ggccactgtg gttgaattgg gagaacccag acatcatgtc agagttactg tttcagaaca
     2581 atgagatcat ctttaaaaat ggggatgatt tacggcaaga tatgctaaca cttcaaatta
     2641 ttcgtattat ggaaaatatc tggcaaaatc aaggtcttga tcttcgaatg ttaccttatg
     2701 gttgtctgtc aatcggtgac tgtgtgggac ttattgaggt ggtgcgaaat tctcacacta
     2761 ttatgcaaat tcagtgcaaa ggcggcttga aaggtgcact gcagttcaac agccacacac
     2821 tacatcagtg gctcaaagac aagaacaaag gagaaatata tgatgcagcc attgacctgt
     2881 ttacacgttc atgtgctgga tactgtgtag ctaccttcat tttgggaatt ggagatcgtc
     2941 acaatagtaa catcatggtg aaagacgatg gacaactgtt tcatatagat tttggacact
     3001 ttttggatca caagaagaaa aaatttggtt ataaacgaga acgtgtgcca tttgttttga
     3061 cacaggattt cttaatagtg attagtaaag gagcccaaga atgcacaaag acaagagaat
     3121 ttgagaggtt tcaggagatg tgttacaagg cttatctagc tattcgacag catgccaatc
     3181 tcttcataaa tcttttctca atgatgcttg gctctggaat gccagaacta caatcttttg
     3241 atgacattgc atacattcga aagaccctag ccttagataa aactgagcaa gaggctttgg
     3301 agtatttcat gaaacaaatg aatgatgcac atcatggtgg ctggacaaca aaaatggatt
     3361 ggatcttcca cacaattaaa cagcatgcat tgaactgatt gccaactttc ttgtacaaag
     3421 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     3481 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     3541 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     3601 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     3661 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3721 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3781 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3841 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3901 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     3961 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     4021 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     4081 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     4141 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     4201 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     4261 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     4321 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     4381 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     4441 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4501 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4561 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     4621 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     4681 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     4741 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     4801 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4861 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4921 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4981 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5041 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5101 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5161 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5221 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5281 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5341 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5401 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5461 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5521 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5581 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5641 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5701 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     5761 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     5821 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     5881 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     5941 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
//
