LOCUS       sequence-320102-        4977 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 4977)
  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 4977)
  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-TBK1
FEATURES             Location/Qualifiers
     source          1..4977
                     /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(2856..2955)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2972..2991)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2973..2991)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2996..3012)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2996..3012)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3443..4234
                     /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      4327..4915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4816..4835
                     /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 agttggcatg cagagcactt ctaatcatct gtggctttta tctgatattt taggccaagg
      721 agctactgca aatgtctttc gtggaagaca taagaaaact ggtgatttat ttgctatcaa
      781 agtatttaat aacataagct tccttcgtcc agtggatgtt caaatgagag aatttgaagt
      841 gttgaaaaaa ctcaatcaca aaaatattgt caaattattt gctattgaag aggagacaac
      901 aacaagacat aaagtactta ttatggaatt ttgtccatgt gggagtttat acactgtttt
      961 agaagaacct tctaatgcct atggactacc agaatctgaa ttcttaattg ttttgcgaga
     1021 tgtggtgggt ggaatgaatc atctacgaga gaatggtata gtgcaccgtg atatcaagcc
     1081 aggaaatatc atgcgtgtta taggggaaga tggacagtct gtgtacaaac tcacagattt
     1141 tggtgcagct agagaattag aagatgatga gcagtttgtt tctctgtatg gcacagaaga
     1201 atatttgcac cctgatatgt atgagagagc agtgctaaga aaagatcatc agaagaaata
     1261 tggagcaaca gttgatcttt ggagcattgg ggtaacattt taccatgcag ctactggatc
     1321 actgccattt agaccctttg aagggcctcg taggaataaa gaagtgatgt ataaaataat
     1381 tacaggaaag ccttctggtg caatatctgg agtacagaaa gcagaaaatg gaccaattga
     1441 ctggagtgga gacatgcctg tttcttgcag tctttctcgg ggtcttcagg ttctacttac
     1501 ccctgttctt gcaaacatcc ttgaagcaga tcaggaaaag tgttggggtt ttgaccagtt
     1561 ttttgcagaa actagtgata tacttcaccg aatggtaatt catgtttttt cgctacaaca
     1621 aatgacagct cataagattt atatacatag ctataatact gctactatat ttcatgaact
     1681 ggtatataaa caaaccaaaa ttatttcttc aaatcaagaa cttatctacg aagggcgacg
     1741 cttagtctta gaacctggaa ggctggcaca acatttccct aaaactactg aggaaaaccc
     1801 tatatttgta gtaagccggg aacctctgga taccatagga ttaatatatg aaaaaatttc
     1861 cctccctaaa gtacatccac gttatgattt agacggggat gctagcatgg ctaaggcaat
     1921 aacaggggtt gtgtgttatg cctgcagaat tgccagtacc ttactgcttt atcaggaatt
     1981 aatgcgaaag gggatacgat ggctgattga attaattaaa gatgattaca atgaaactgt
     2041 tcacaaaaag acagaagttg tgatcacatt ggatttctgt atcagaaaca ttgaaaaaac
     2101 tgtgaaagta tatgaaaagt tgatgaagat caacctggaa gcggcagagt taggtgaaat
     2161 ttcagacata cacaccaaat tgttgagact ttccagttct cagggaacaa tagaaaccag
     2221 tcttcaggat atcgacagca gattatctcc aggtggatca ctggcagacg catgggcaca
     2281 tcaagaaggc actcatccga aagacagaaa tgtagaaaaa ctacaagtcc tgttaaattg
     2341 catgacagag atttactatc agttcaaaaa agaccaagca gaacgtagat tagcttataa
     2401 tgaagaacaa atccacaaat ttgataagca aaaactgtat taccatgcca caaaagctat
     2461 gacgcacttt acagatgaat gtgttaaaaa gtatgaggca tttttgaata agtcagaaga
     2521 atggataaga aagatgcttc atcttaggaa acagttatta tcgctgacta atcagtgttt
     2581 tgatattgaa gaagaagtat caaaatatca agaatatact aatgagttac aagaaactct
     2641 gcctcagaaa atgtttacag cttccagtgg aatcaaacat accatgaccc caatttatcc
     2701 aagttctaac acattagtag aaatgactct tggtatgaag aaattaaagg aagagatgga
     2761 aggggtggtt aaagaacttg ctgaaaataa ccacatttta gaaaggtttg gctctttaac
     2821 catggatggt ggccttcgca acgttgactg tctttaccca actttcttgt acaaagttgg
     2881 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2941 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     3001 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     3061 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
     3121 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     3181 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     3241 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     3301 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     3361 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     3421 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     3481 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     3541 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     3601 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     3661 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     3721 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     3781 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     3841 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     3901 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     3961 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     4021 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     4081 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     4141 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     4201 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     4261 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4321 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4381 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4441 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4501 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4561 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4621 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4681 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4741 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4801 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4861 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4921 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
