LOCUS       sequence-496541-        5997 bp    DNA     circular SYN 08-JAN-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5997)
  AUTHORS   Johannessen CM, Boehm JS, Kim SY, Thomas SR, Wardwell L, Johnson LA,
            Emery CM, Stransky N, Cogdill AP, Barretina J, Caponigro G, 
            Hieronymus H, Murray RR, Salehi-Ashtiani K, Hill DE, Vidal M, Zhao 
            JJ, Yang X, Alkan O, Kim S, Harris JL, Wilson CJ, Myer VE, Finan PM,
            Root DE, Roberts TM, Golub T, Flaherty KT, Dummer R, Weber BL, 
            Sellers WR, Schlegel R, Wargo JA, Hahn WC, Garraway LA
  TITLE     COT drives resistance to RAF inhibition through MAP kinase pathway 
            reactivation.
  JOURNAL   Nature. 2010 Dec 16;468(7326):968-72. doi: 10.1038/nature09627
  PUBMED    21107320
REFERENCE   2  (bases 1 to 5997)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 8, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2010-12-16- 16"; volume: "468"; issue: "7326"; 
            pages: "968-72"
COMMENT     Sequence Label: pDONR223-PIK3CA
FEATURES             Location/Qualifiers
     source          1..5997
                     /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(3876..3975)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3992..4011)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3993..4011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4016..4032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4016..4032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4463..5254
                     /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      5347..5935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5836..5855
                     /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 aggttttgga
     3781 gtatttcatg aaacaaatga atgatgcaca tcatggtggc tggacaacaa aaatggattg
     3841 gatcttccac acaattaaac agcatgcatt gaacttgcca actttcttgt acaaagttgg
     3901 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     3961 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     4021 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     4081 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
     4141 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     4201 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     4261 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     4321 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     4381 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     4441 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     4501 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     4561 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     4621 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     4681 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     4741 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     4801 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     4861 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     4921 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     4981 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     5041 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     5101 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     5161 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     5221 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     5281 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
