LOCUS       sequence-424530-        5460 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 5460)
  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 5460)
  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-TYRO3
FEATURES             Location/Qualifiers
     source          1..5460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..87)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(70..86)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(79..101)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      150..236
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      328..355
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(514..531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(685..1273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(765..784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1366..2157)
                     /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"
     primer_bind     2588..2604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2588..2604
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2609..2628
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2609..2627
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2645..2744
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(join(5415..5460,1..54))
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
       61 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt
      121 taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
      181 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
      241 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
      301 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
      361 ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cactgcccgc
      421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1201 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1321 cgcgtaactc acgttaaggg attttggtca tgggtggctc gagggttatt tgccgactac
     1381 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     1441 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     1501 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     1561 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     1621 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     1681 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     1741 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     1801 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     1861 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     1921 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     1981 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     2041 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     2101 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     2161 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     2221 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     2281 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     2341 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     2401 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     2461 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg tctagaggag
     2521 gcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca
     2581 gagctgccag gaaacagcta tgaccatgta atacgactca ctatagggga tatcagctgg
     2641 atggcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa
     2701 gcaatgcttt cttataatgc caactttgta caagaaagtt gggtaacagc tactgtgtgg
     2761 cagtagccct tgctgcagca gcaaaagcct ctgtgtctca ttgaggggac tctctggctg
     2821 gtgctctgcc tgccctggct gctcagccag ccctccgggg gtgagtatgt accgacagtc
     2881 actgggagtg ccacccactg cccccatgcc actgccatcc ccagccccac tgtagggctg
     2941 atccccgcca ggtagctcca tgctgcctcc cgcagtgggc tcctcagctc tctcgatgtt
     3001 gatgtataag gggtcctggc tggcagatag cacagacagc tggcccaaga tgttctccag
     3061 ttccattcgc agacaagtaa agctcgggcg ctgcttgggg tcagcactcc agcactggta
     3121 catgagatca tacacgtcct ccatacactc cggaggctgt ttcaggcggt tcccgccaat
     3181 gaggtagttg taaatctcag cgttttcgat gccagcatat ggcgtctgcc cacgtgtcat
     3241 gatctcccac atggtcaccc cgaacgccca cacgtcactc tgcacagtat acaggttgtc
     3301 ggccaggctc tccagggcca gccacttgac aggcagtttg gaggcacagc cttgacgata
     3361 gtagtcccca ctgtagatct tccgggagag tccgaagtca gccacacaca ctgtcatgtc
     3421 ctctgccagc atgcaattcc gagcagccag gtctcggtgg atgaagttcc gagagctcag
     3481 gtactccatg ccgcaggcaa tgtccaccat gaaccggatc agggtctgga ggggtaggtt
     3541 aaaggggttc tccccaatcc gggaggcgag caggaaggca tgcaggtccc catgcttcat
     3601 gaagggcaag atgaccatgg ggatggggag acggccttta gccctgctcc ggaggcttac
     3661 cccaacaagt ttggccacgt gtggatggtc aaactccttc atgcaagctg cttccctgag
     3721 gaactcttca atgtcgcttg aggcaatgat gtcagctttc agcatcttca cagccacttt
     3781 cacaaaggag ctatcctctt gcttcagctg ggcctcccgc actgaaccaa actctccttt
     3841 gcccaacatc cggcccaggg tgaactgctg ctctgggatg agcacatcct ccagtttttc
     3901 ctttagttca tcgctgatgc ccaagctgtc caatgtggcc tcgatgcgct cgggcctttc
     3961 tcgattgaag gaccgggctg cccggaagtg aacggctggc tctccccggg ccatgacact
     4021 gtcaaaggct tgcccaaacc gcgtctcttt ccgtctcttt cgaagcagga tgagggccag
     4081 ggcagcagcc gtcaccaggg ccgttagcac accaaggacc acaggtaccc aggatgtgcg
     4141 gctgtgagga gggccctgct ggcctgcacg gtcatgagaa gagaccacca gtggctgact
     4201 ccagggtcca cagccaactg cattggagac gcacacacgt acgatcaggt ccttttgggg
     4261 atcccagcct gtcaaattgg ccctggtccc ctccactgtc agctcatcct gggttccatt
     4321 gtcttgaacc caggacagtt tgtagggtcc cagggggcct tccaaagggg cctcggggat
     4381 cacttcttcc cactccaaga tgaggcctga atctgtgcgg atggcatgga ggttttgggg
     4441 agcgctggct ggggctagac ccttggtctg aaagggcacc cagtcagcat agggagaggg
     4501 ccccaaggca ttggcacagc gcaccctgag gctgtagttg gtggcaggca ccaggtcccg
     4561 gagcaggcag gtaaaggggg gcacagggac cacaacagcc aggacttccc agcctcctgg
     4621 ggcctgtgtc acctgaactg tacaggactg tagcagagct cggccatcgg cacctggcat
     4681 ccaggccaca ctagcgttgc tgctggaaag ctttgtcacg gtgatgttga agggggctgc
     4741 aggcagtgct tgaaggtgaa cagtggctgt gcgagaagag gccaggcctt ttaggttgtg
     4801 agcttcacag gaaaacatgg tgctctgggt cacccctgtt acatttaaaa cagatggaga
     4861 gggagcgggt cccccgatct tcgtagttcc tctccaccag acaatggtaa caggttcagg
     4921 gggacccaca gcctcacaag acagttggaa aggggcattg ggtggcactg ccagatcttt
     4981 tggctccact gtgaaaaatg gcacaccttc taccgtgagc cacactggct gggagatctc
     5041 ggtttcaccc ccatcctcca cctggcacca gtaccggccg gcgtcagagc gctccactga
     5101 cttcaggctg aggaagccga tccagtgctg ctcgctgact gggatgtaca actggtccaa
     5161 gttctggacc acagccccat ccttcaccca ctggatgtca ggctcctcca tcccctccac
     5221 actgcagttg agcttcaccg gctgcccctg agacactgtc agcttcaccg gggctcccat
     5281 gagcttcaga cctgcggcgg cggactccgg gagcagcaga gaagccagag ccgccagcag
     5341 cagcccgagc cgcagtggcg gcggcagcgg cagcggcggg agccccggcc gccccatgct
     5401 ccgcctcagc gccatgccaa cttttttgta caaagttggc attataaaaa agcattgctc
//
