LOCUS       sequence-162444-        5999 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 5999)
  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 5999)
  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.L699M
FEATURES             Location/Qualifiers
     source          1..5999
                     /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(3399..3498)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3515..3534)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3516..3534)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3539..3555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3539..3555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3986..4777
                     /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      4870..5458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5359..5378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5612..5629
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5788..5815)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5907..5993)
                     /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 atgcctccac gaccatcatc aggtgaactg tggggcatcc acttgatgcc
      241 cccaagaatc ctagtagaat gtttactacc aaatggaatg atagtgactt tagaatgcct
      301 ccgtgaggct acattaataa ccataaagca tgaactattt aaagaagcaa gaaaataccc
      361 cctccatcaa cttcttcaag atgaatcttc ttacattttc gtaagtgtta ctcaagaagc
      421 agaaagggaa gaattttttg atgaaacaag acgactttgt gaccttcggc tttttcaacc
      481 ctttttaaaa gtaattgaac cagtaggcaa ccgtgaagaa aagatcctca atcgagaaat
      541 tggttttgct atcggcatgc cagtgtgtga atttgatatg gttaaagatc cagaagtaca
      601 ggacttccga agaaatattc tgaacgtttg taaagaagct gtggatctta gggacctcaa
      661 ttcacctcat agtagagcaa tgtatgtcta tcctccaaat gtagaatctt caccagaatt
      721 gccaaagcac atatataata aattagataa agggcaaata atagtggtga tctgggtaat
      781 agtttctcca aataatgaca agcagaagta tactctgaaa atcaaccatg actgtgtacc
      841 agaacaagta attgctgaag caatcaggaa aaaaactcga agtatgttgc tatcctctga
      901 acaactaaaa ctctgtgttt tagaatatca gggcaagtat attttaaaag tgtgtggatg
      961 tgatgaatac ttcctagaaa aatatcctct gagtcagtat aagtatataa gaagctgtat
     1021 aatgcttggg aggatgccca atttgatgtt gatggctaaa gaaagccttt attctcaact
     1081 gccaatggac tgttttacaa tgccatctta ttccagacgc atttccacag ctacaccata
     1141 tatgaatgga gaaacatcta caaaatccct ttgggttata aatagtgcac tcagaataaa
     1201 aattctttgt gcaacctacg tgaatgtaaa tattcgagac attgataaga tctatgttcg
     1261 aacaggtatc taccatggag gagaaccctt atgtgacaat gtgaacactc aaagagtacc
     1321 ttgttccaat cccaggtgga atgaatggct gaattatgat atatacattc ctgatcttcc
     1381 tcgtgctgct cgactttgcc tttccatttg ctctgttaaa ggccgaaagg gtgctaaaga
     1441 ggaacactgt ccattggcat ggggaaatat aaacttgttt gattacacag acactctagt
     1501 atctggaaaa atggctttga atctttggcc agtacctcat ggattagaag atttgctgaa
     1561 ccctattggt gttactggat caaatccaaa taaagaaact ccatgcttag agttggagtt
     1621 tgactggttc agcagtgtgg taaagttccc agatatgtca gtgattgaag agcatgccaa
     1681 ttggtctgta tcccgagaag caggatttag ctattcccac gcaggactga gtaacagact
     1741 agctagagac aatgaattaa gggaaaatga caaagaacag ctcaaagcaa tttctacacg
     1801 agatcctctc tctgaaatca ctgagcagga gaaagatttt ctatggagtc acagacacta
     1861 ttgtgtaact atccccgaaa ttctacccaa attgcttctg tctgttaaat ggaattctag
     1921 agatgaagta gcccagatgt attgcttggt aaaagattgg cctccaatca aacctgaaca
     1981 ggctatggaa cttctggact gtaattaccc agatcctatg gttcgaggtt ttgctgttcg
     2041 gtgcttggaa aaatatttaa cagatgacaa actttctcag tatttaattc agctagtaca
     2101 ggtcctaaaa tatgaacaat atttggataa cttgcttgtg agatttttac tgaagaaagc
     2161 attgactaat caaaggattg ggcacttttt cttttggcat ttaaaatctg agatgcacaa
     2221 taaaacagtt agccagaggt ttggcctgct tttggagtcc tattgtcgtg catgtgggat
     2281 gtatatgaag cacctgaata ggcaagtcga ggcaatggaa aagctcatta acttaactga
     2341 cattctcaaa caggagaaga aggatgaaac acaaaaggta cagatgaagt ttttagttga
     2401 gcaaatgagg cgaccagatt tcatggatgc tctacagggc tttctgtctc ctctaaaccc
     2461 tgctcatcaa ctaggaaacc tcaggcttga agagtgtcga attatgtcct ctgcaaaaag
     2521 gccactgtgg ttgaattggg agaacccaga catcatgtca gagttactgt ttcagaacaa
     2581 tgagatcatc tttaaaaatg gggatgattt acggcaagat atgctaacac ttcaaattat
     2641 tcgtattatg gaaaatatct ggcaaaatca aggtcttgat cttcgaatgt taccttatgg
     2701 ttgtctgtca atcggtgact gtgtgggact tattgaggtg gtgcgaaatt ctcacactat
     2761 tatgcaaatt cagtgcaaag gcggcttgaa aggtgcactg cagttcaaca gccacacact
     2821 acatcagtgg ctcaaagaca agaacaaagg agaaatatat gatgcagcca ttgacctgtt
     2881 tacacgttca tgtgctggat actgtgtagc taccttcatt ttgggaattg gagatcgtca
     2941 caatagtaac atcatggtga aagacgatgg acaactgttt catatagatt ttggacactt
     3001 tttggatcac aagaagaaaa aatttggtta taaacgagaa cgtgtgccat ttgttttgac
     3061 acaggatttc ttaatagtga ttagtaaagg agcccaagaa tgcacaaaga caagagaatt
     3121 tgagaggttt caggagatgt gttacaaggc ttatctagct attcgacagc atgccaatct
     3181 cttcataaat cttttctcaa tgatgcttgg ctctggaatg ccagaactac aatcttttga
     3241 tgacattgca tacattcgaa agaccctagc cttagataaa actgagcaag aggctttgga
     3301 gtatttcatg aaacaaatga atgatgcaca tcatggtggc tggacaacaa aaatggattg
     3361 gatcttccac acaattaaac agcatgcatt gaactgattg ccaactttct tgtacaaagt
     3421 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     3481 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     3541 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     3601 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
     3661 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     3721 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     3781 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     3841 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     3901 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     3961 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
     4021 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     4081 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     4141 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     4201 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     4261 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     4321 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     4381 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     4441 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     4501 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     4561 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     4621 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     4681 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     4741 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     4801 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5521 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5581 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     5641 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     5701 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
     5761 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     5821 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     5881 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     5941 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcct
//
