LOCUS       sequence-344993-        4083 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 4083)
  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 4083)
  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-SGK
FEATURES             Location/Qualifiers
     source          1..4083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1426..1525)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1542..1561)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1543..1561)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1566..1582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1566..1582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2013..2804
                     /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      2897..3485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3386..3405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3639..3656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3815..3842)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3934..4020)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4069..4083,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4083,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg catgacggtg aaaactgagg ctgctaaggg caccctcact tactccagga
      181 tgaggggcat ggtggcaatt ctcatcgctt tcatgaagca gaggaggatg ggtctgaacg
      241 actttattca gaagattgcc aataactcct atgcatgcaa acaccctgaa gttcagtcca
      301 tcttgaagat ctcccaacct caggagcctg agcttatgaa tgccaaccct tctcctccac
      361 caagtccttc tcagcaaatc aaccttggcc cgtcgtccaa tcctcatgct aaaccatctg
      421 actttcactt cttgaaagtg atcggaaagg gcagttttgg aaaggttctt ctagcaagac
      481 acaaggcaga agaagtgttc tatgcagtca aagttttaca gaagaaagca atcctgaaaa
      541 agaaagagga gaagcatatt atgtcggagc ggaatgttct gttgaagaat gtgaagcacc
      601 ctttcctggt gggccttcac ttctctttcc agactgctga caaattgtac tttgtcctag
      661 actacattaa tggtggagag ttgttctacc atctccagag ggaacgctgc ttcctggaac
      721 cacgggctcg tttctatgct gctgaaatag ccagtgcctt gggctacctg cattcactga
      781 acatcgttta tagagactta aaaccagaga atattttgct agattcacag ggacacattg
      841 tccttactga cttcggactc tgcaaggaga acattgaaca caacagcaca acatccacct
      901 tctgtggcac gccggagtat ctcgcacctg aggtgcttca taagcagcct tatgacagga
      961 ctgtggactg gtggtgcctg ggagctgtct tgtatgagat gctgtatggc ctgccgcctt
     1021 tttatagccg aaacacagct gaaatgtacg acaacattct gaacaagcct ctccagctga
     1081 aaccaaatat tacaaattcc gcaagacacc tcctggaggg cctcctgcag aaggacagga
     1141 caaagcggct cggggccaag gatgacttca tggagattaa gagtcatgtc ttcttctcct
     1201 taattaactg ggatgatctc attaataaga agattactcc cccttttaac ccaaatgtga
     1261 gtgggcccaa cgacctacgg cactttgacc ccgagtttac cgaagagcct gtccccaact
     1321 ccattggcaa gtcccctgac agcgtcctcg tcacagccag cgtcaaggaa gctgccgagg
     1381 ctttcctagg cttttcctat gcgcctccca cggactcttt actctgccca actttcttgt
     1441 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1501 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1561 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1621 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     1681 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     1741 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     1801 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     1861 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     1921 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     1981 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     2041 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     2101 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     2161 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     2221 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     2281 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     2341 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     2401 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     2461 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     2521 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     2581 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     2641 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     2701 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     2761 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     2821 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     2881 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2941 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3001 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3061 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3121 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3181 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3241 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3301 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3361 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3421 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3481 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3541 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3601 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3661 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3721 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     3781 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     3841 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     3901 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     3961 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     4021 atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac
     4081 gtt
//
