LOCUS       sequence-443283-        6867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6867)
  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 6867)
  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"; journalName: 
            "Nature"; date: "2010-12-16- 16"; volume: "468"; issue: "7326"; 
            pages: "968-72"
COMMENT     Sequence Label: pDONR223-PIK3R4
FEATURES             Location/Qualifiers
     source          1..6867
                     /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(4746..4845)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4862..4881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4863..4881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4886..4902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4886..4902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5333..6124
                     /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      6217..6805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6706..6725
                     /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 atgggaaatc agcttgctgg cattgctccc tcccagatcc tttctgtaga
      721 gagttatttt tcagatattc atgactttga atatgataaa agcctgggga gtactcggtt
      781 ttttaaagtt gctcgagcca agcaccgaga aggcctggtc gttgtgaagg tttttgcaat
      841 tcaggatccc acattgcctt taaccagcta taaacaagag ctggaggaac tgaaaatcag
      901 gcttaattct gcacagaatt gtctaccttt ccagaaagca tcagaaaaag catctgagaa
      961 agcagctatg ctctttaggc agtatgtgcg agacaatctc tatgatcgca tcagtacccg
     1021 tccattcttg aataacattg agaagcgctg gattgctttc cagatcctga cagctgtgga
     1081 ccaagcacac aaatctggag ttcgtcatgg ggacatcaag actgagaatg tgatggtcac
     1141 cagttggaat tgggttcttc taactgattt tgccagtttt aagcccactt atcttccaga
     1201 agacaacccg gcagatttca attatttctt tgacacatca cggaggagaa cttgctatat
     1261 tgctcctgaa cgttttgttg atggtgggat gtttgccact gagttagaat atatgagaga
     1321 tccttcaact ccgcttgtag acttaaatag caatcagaga acaagaggag agttgaagag
     1381 agcaatggac atcttttcag caggttgtgt gatagctgag ctttttacag aaggtgtacc
     1441 attatttgat ctctctcaac ttttggctta tagaaatgga cattttttcc ctgaacaagt
     1501 gctaaataaa attgaagatc acagtatcag agaattggta actcagatga ttcaccgtga
     1561 gccagataaa cgtttagagg cagaagatta cttaaaacag cagcgtggca atgcctttcc
     1621 tgaaatattt tacacttttc ttcagcccta catggcccag tttgccaagg aaacgtttct
     1681 ttctgcagat gagcgtattc tggttatacg gaaggatttg ggcaacatta ttcacaatct
     1741 ctgtggacat gatctgccag aaaaagccga aggagagcct aaggaaaatg ggctggttat
     1801 cttggtatct gttataacat cctgcctaca gacccttaaa tactgtgatt ccaaactagc
     1861 tgctttggaa ctgattcttc atttggctcc aagattaagt gttgaaatcc ttttggatcg
     1921 tattactcca tatcttttgc atttcagcaa tgactctgtt cctagggtga gggctgaagc
     1981 cttgaggacg ttgaccaaag ttcttgctct cgtcaaagag gttcctcgta atgatatcaa
     2041 tatttatccg gaatacattc tgccaggcat agcccactta gcccaagatg atgctactat
     2101 cgttagacta gcctatgctg aaaacatagc tctgctggca gaaacagctc tgagattcct
     2161 ggaattagta cagttaaaaa atctcaatat ggaaaatgac cccaataatg aagaaataga
     2221 tgaggttaca catccaaatg gaaattatga cacagagctc caagccttac atgaaatggt
     2281 ccagcagaaa gttgttactt tgctaagtga ccctgaaaat attgtaaaac aaaccttgat
     2341 ggaaaatgga ataacacggc tgtgtgtatt ctttggacgt cagaaagcca acgatgtttt
     2401 gttgtcccac atgattactt tcctaaatga taagaatgat tggcatctac gtggagcatt
     2461 ttttgatagt atagttggtg ttgctgccta tgttggctgg caaagctcct caattctcaa
     2521 gcctctgctg caacaaggtc ttagtgatgc tgaggaattt gtcattgtga aagctcttta
     2581 tgcccttact tgtatgtgcc agttaggact gctacaaaaa ccccatgttt acgaatttgc
     2641 cagtgatatt gcccccttcc tgtgtcatcc caatttatgg atacgttatg gtgccgtggg
     2701 atttatcaca gtggtagctc gtcaaataag tacagctgat gtctactgta aactgatgcc
     2761 ttatcttgac ccatatatta cccaaccaat aatacagatt gaaagaaaac ttgttctgct
     2821 cagtgtttta aaggaaccag taagtcgttc tatatttgat tatgctttga ggtctaaaga
     2881 tattactagc ttgttcagac atcttcacat gcgtcagaag aaacgaaatg gttctcttcc
     2941 cgactgccct ccgccagagg atcctgccat agcacagctt ctgaagaagt tgctctcaca
     3001 gggaatgaca gaggaagagg aagacaaact tctggcactg aaagacttca tgatgaaatc
     3061 taataaagca aaggccaata tagtggacca gagccatctt catgatagta gtcagaaagg
     3121 tgtaattgac ttggcagctt taggcataac tgggagacaa gttgatcttg ttaaaaccaa
     3181 acaagaacca gatgacaaac gggccagaaa acatgtaaaa caagactcaa atgtaaatga
     3241 agaatggaaa agcatgtttg ggtcactgga cccaccaaac atgccacagg ccctacctaa
     3301 agggagtgat caggaggtga ttcagactgg gaaacctcct cgttccgagt cctctgctgg
     3361 catttgtgtc cctttgtcaa cttcttcaca ggttccagaa gtgacaactg tccaaaataa
     3421 aaaaccagta ataccggttt taagtagtac aatcttacca tccacctatc agattcgaat
     3481 tacaacttgt aaaactgaac ttcagcaact catccagcaa aagcgggagc agtgcaatgc
     3541 tgagagaata gctaagcaga tgatggaaaa tgctgaatgg gagagtaaac caccaccacc
     3601 tggatggcgt cctaaagggc tgttagttgc ccatcttcat gagcataaat ctgctgtgaa
     3661 tcgaattaga gtctctgatg aacactcact ttttgcaaca tgttcaaatg atggcacagt
     3721 gaaaatctgg aacagtcaaa agatggaggg gaagaccacc actaccagat ctattcttac
     3781 atacagccga attggaggac gagtcaagac gctcacattc tgccaaggct cccactattt
     3841 agccatagca tctgataatg gtgctgtcca gcttcttgga attgaggctt ctaagctgcc
     3901 caagtctcct aaaatccatc ctctacaaag cagaattcta gatcagaagg aggacggttg
     3961 tgttgtggat atgcatcact tcaactctgg agcacagtct gttcttgcct atgccactgt
     4021 gaatggctct ctggttggct gggaccttag gtcttcaagc aatgcgtgga ctttaaagca
     4081 tgatttaaag tcgggcctca tcacttcctt tgctgtggac atccaccaat gctggctctg
     4141 cattggtaca agcagtggta ccatggcttg ttgggacatg aggttccagt tgccaatttc
     4201 aagtcactgt catccttcca gggctcgaat cagacgcctc tcaatgcacc ctctgtatca
     4261 gtcctgggtg attgcagctg ttcagggcaa caacgaagtg tccatgtggg acatggagac
     4321 tggtgacaga agatttactc tctgggccag cagtgcacca ccactttctg aattacagcc
     4381 ttctcctcat agcgtccatg gtatctactg tagtcctgca gatggaaatc ctatcctact
     4441 aacagctggc tcagatatga aaataaggtt ttgggacttg gcttacccag aaaggtccta
     4501 tgttgttgca ggaagtacta gttccccatc tgtgtcctac tacaggaaaa taattgaagg
     4561 cactgaagtt gtccaggaaa ttcagaataa gcagaaagta ggaccaagtg atgacacccc
     4621 tcgaaggggc ccagagtccc tgcccgtggg acatcatgac atcatcactg atgtcgccac
     4681 attccagacc acacagggct tcatcgtaac tgcttctaga gatgggattg tgaaggtgtg
     4741 gaaattgcca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4801 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     4861 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     4921 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac
     4981 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     5041 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     5101 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     5161 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     5221 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     5281 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     5341 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     5401 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     5461 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     5521 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     5581 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     5641 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     5701 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     5761 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     5821 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     5881 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     5941 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     6001 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     6061 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     6121 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     6181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6841 cttttgctgg ccttttgctc acatgtt
//
