LOCUS       sequence-162306-        6000 bp    DNA     circular SYN 14-APR-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 Apr 14, 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.H1047R
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3879..3978)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3995..4014)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3996..4014)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4019..4035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4019..4035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4466..5257
                     /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      5350..5938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5839..5858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgcctccac gaccatcatc aggtgaactg tggggcatcc acttgatgcc
      721 cccaagaatc ctagtagaat gtttactacc aaatggaatg atagtgactt tagaatgcct
      781 ccgtgaggct acattaataa ccataaagca tgaactattt aaagaagcaa gaaaataccc
      841 cctccatcaa cttcttcaag atgaatcttc ttacattttc gtaagtgtta ctcaagaagc
      901 agaaagggaa gaattttttg atgaaacaag acgactttgt gaccttcggc tttttcaacc
      961 ctttttaaaa gtaattgaac cagtaggcaa ccgtgaagaa aagatcctca atcgagaaat
     1021 tggttttgct atcggcatgc cagtgtgtga atttgatatg gttaaagatc cagaagtaca
     1081 ggacttccga agaaatattc tgaacgtttg taaagaagct gtggatctta gggacctcaa
     1141 ttcacctcat agtagagcaa tgtatgtcta tcctccaaat gtagaatctt caccagaatt
     1201 gccaaagcac atatataata aattagataa agggcaaata atagtggtga tctgggtaat
     1261 agtttctcca aataatgaca agcagaagta tactctgaaa atcaaccatg actgtgtacc
     1321 agaacaagta attgctgaag caatcaggaa aaaaactcga agtatgttgc tatcctctga
     1381 acaactaaaa ctctgtgttt tagaatatca gggcaagtat attttaaaag tgtgtggatg
     1441 tgatgaatac ttcctagaaa aatatcctct gagtcagtat aagtatataa gaagctgtat
     1501 aatgcttggg aggatgccca atttgatgtt gatggctaaa gaaagccttt attctcaact
     1561 gccaatggac tgttttacaa tgccatctta ttccagacgc atttccacag ctacaccata
     1621 tatgaatgga gaaacatcta caaaatccct ttgggttata aatagtgcac tcagaataaa
     1681 aattctttgt gcaacctacg tgaatgtaaa tattcgagac attgataaga tctatgttcg
     1741 aacaggtatc taccatggag gagaaccctt atgtgacaat gtgaacactc aaagagtacc
     1801 ttgttccaat cccaggtgga atgaatggct gaattatgat atatacattc ctgatcttcc
     1861 tcgtgctgct cgactttgcc tttccatttg ctctgttaaa ggccgaaagg gtgctaaaga
     1921 ggaacactgt ccattggcat ggggaaatat aaacttgttt gattacacag acactctagt
     1981 atctggaaaa atggctttga atctttggcc agtacctcat ggattagaag atttgctgaa
     2041 ccctattggt gttactggat caaatccaaa taaagaaact ccatgcttag agttggagtt
     2101 tgactggttc agcagtgtgg taaagttccc agatatgtca gtgattgaag agcatgccaa
     2161 ttggtctgta tcccgagaag caggatttag ctattcccac gcaggactga gtaacagact
     2221 agctagagac aatgaattaa gggaaaatga caaagaacag ctcaaagcaa tttctacacg
     2281 agatcctctc tctgaaatca ctgagcagga gaaagatttt ctatggagtc acagacacta
     2341 ttgtgtaact atccccgaaa ttctacccaa attgcttctg tctgttaaat ggaattctag
     2401 agatgaagta gcccagatgt attgcttggt aaaagattgg cctccaatca aacctgaaca
     2461 ggctatggaa cttctggact gtaattaccc agatcctatg gttcgaggtt ttgctgttcg
     2521 gtgcttggaa aaatatttaa cagatgacaa actttctcag tatttaattc agctagtaca
     2581 ggtcctaaaa tatgaacaat atttggataa cttgcttgtg agatttttac tgaagaaagc
     2641 attgactaat caaaggattg ggcacttttt cttttggcat ttaaaatctg agatgcacaa
     2701 taaaacagtt agccagaggt ttggcctgct tttggagtcc tattgtcgtg catgtgggat
     2761 gtatttgaag cacctgaata ggcaagtcga ggcaatggaa aagctcatta acttaactga
     2821 cattctcaaa caggagaaga aggatgaaac acaaaaggta cagatgaagt ttttagttga
     2881 gcaaatgagg cgaccagatt tcatggatgc tctacagggc tttctgtctc ctctaaaccc
     2941 tgctcatcaa ctaggaaacc tcaggcttga agagtgtcga attatgtcct ctgcaaaaag
     3001 gccactgtgg ttgaattggg agaacccaga catcatgtca gagttactgt ttcagaacaa
     3061 tgagatcatc tttaaaaatg gggatgattt acggcaagat atgctaacac ttcaaattat
     3121 tcgtattatg gaaaatatct ggcaaaatca aggtcttgat cttcgaatgt taccttatgg
     3181 ttgtctgtca atcggtgact gtgtgggact tattgaggtg gtgcgaaatt ctcacactat
     3241 tatgcaaatt cagtgcaaag gcggcttgaa aggtgcactg cagttcaaca gccacacact
     3301 acatcagtgg ctcaaagaca agaacaaagg agaaatatat gatgcagcca ttgacctgtt
     3361 tacacgttca tgtgctggat actgtgtagc taccttcatt ttgggaattg gagatcgtca
     3421 caatagtaac atcatggtga aagacgatgg acaactgttt catatagatt ttggacactt
     3481 tttggatcac aagaagaaaa aatttggtta taaacgagaa cgtgtgccat ttgttttgac
     3541 acaggatttc ttaatagtga ttagtaaagg agcccaagaa tgcacaaaga caagagaatt
     3601 tgagaggttt caggagatgt gttacaaggc ttatctagct attcgacagc atgccaatct
     3661 cttcataaat cttttctcaa tgatgcttgg ctctggaatg ccagaactac aatcttttga
     3721 tgacattgca tacattcgaa agaccctagc cttagataaa actgagcaag aggctttgga
     3781 gtatttcatg aaacaaatga atgatgcacg tcatggtggc tggacaacaa aaatggattg
     3841 gatcttccac acaattaaac agcatgcatt gaactgattg ccaactttct tgtacaaagt
     3901 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     3961 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     4021 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     4081 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
     4141 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     4201 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     4261 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     4321 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     4381 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     4441 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
     4501 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     4561 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     4621 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     4681 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     4741 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     4801 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     4861 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     4921 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     4981 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     5041 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     5101 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     5161 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     5221 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     5281 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5341 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5401 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5461 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5521 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5581 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5641 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5701 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5761 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5821 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5881 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5941 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
