LOCUS       sequence-267449-        4965 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 4965)
  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 4965)
  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-EEF2K
FEATURES             Location/Qualifiers
     source          1..4965
                     /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(2308..2407)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2424..2443)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2425..2443)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2448..2464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2448..2464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2895..3686
                     /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      3779..4367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4268..4287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4521..4538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4697..4724)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4816..4902)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4951..4965,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4965,1..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 catggcagac gaagatctca tcttccgcct ggaaggcgtt gatggcggcc
      181 agtccccccg agctggccgt gatggtgatt ctgatgggga cagcgacgat gaggaaggtt
      241 acttcatctg ccccatcacg gatgacccaa gctcgaacca gaatgtcaat tccaaggtta
      301 ataagtacta cagcaaccta acaaaaagtg agcggtatag ctccagcggg tccccggcaa
      361 actccttcca cttcaaggaa gcctggaagc acgcaatcca gaaggccaag cacatgcccg
      421 acccctgggc tgagttccac ctggaagata ttgccaccga acgtgctact cgacacaggt
      481 acaacgccgt caccggggaa tggctggatg atgaagttct gatcaagatg gcatctcagc
      541 ccttcggccg aggagcaatg agggagtgct tccggacgaa gaagctctcc aacttcttgc
      601 atgcccagca gtggaaggga gcctccaact acgtggcgaa gcgctacatc gagcccgtag
      661 accgggatgt gtactttgag gacgtgcgtc tacagatgga ggccaagctc tggggggagg
      721 agtataatcg gcacaagccc cccaagcagg tggacatcat gcagatgtgc atcatcgagc
      781 tgaaggacag accgggcaag cccctcttcc acctggagca ctacatcgag ggcaagtaca
      841 tcaagtacaa ctccaactct ggctttgtcc gcgatgacaa catccgcctg acgccgcagg
      901 ccttcagcca cttcactttt gagcgttccg gccatcagct gatagtggtg gacatccagg
      961 gagttgggga tctctacact gacccacaga tccacacgga gacgggcact gactttggag
     1021 acggcaacct aggtgtccgc gggatggcgc tcttcttcta ctctcatgcc tgcaaccgga
     1081 tttgcgagag catgggcctt gctccctttg acctctcgcc ccgggagagg gatgcagtga
     1141 atcagaacac caagctgctg caatcagcca agaccatctt gagaggaaca gaggaaaaat
     1201 gtgggagccc ccgagtaagg accctctctg ggagccggcc acccctgctc cgtccccttt
     1261 cagagaactc tggagacgag aacatgagcg acgtgacctt cgactctctc ccttcttccc
     1321 catcttcggc cacaccacac agccagaagc tagaccacct ccattggcca gtgttcagtg
     1381 acctcgataa catggcatcc agagaccatg atcatctaga caaccaccgg gagtctgaga
     1441 atagtgggga cagcggatac cccagtgaga agcggggtga gctggatgac cctgagcccc
     1501 gagaacatgg ccactcatac agtaatcgga agtacgagtc tgacgaagac agcctgggca
     1561 gctctggacg ggtatgtgta gagaagtgga atctcctcaa ctcctcccgc ctccacctgc
     1621 cgagggcttc ggccgtggcc ctggaagtgc aaaggcttaa tgctctggac ctcgaaaaga
     1681 aaatcgggaa gtccattttg gggaaggtcc atctggccat ggtgcgctac cacgagggtg
     1741 ggcgcttctg cgagaagggc gaggagtggg accaggagtc ggctgtcttc cacctggagc
     1801 acgcagccaa cctgggcgag ctggaggcca tcgtgggcct gggactcatg tactcgcagt
     1861 tgcctcatca catcctagcc gatgtctctc tgaaggagac agaagagaac aaaaccaaag
     1921 gatttgatta cttactaaag gccgctgaag ctggcgacag gcagtccatg atcctagtgg
     1981 cgcgagcttt tgactctggc cagaacctca gcccggacag gtgccaagac tggctagagg
     2041 ccctgcactg gtacaacact gccctggaga tgacggactg tgatgagggc ggtgagtacg
     2101 acggaatgca ggacgagccc cggtacatga tgctggccag ggaggccgag atgctgttca
     2161 caggaggcta cgggctggag aaggacccgc agagatcagg ggacttgtat acccaggcag
     2221 cagaggcagc gatggaagcc atgaagggcc gactggccaa ccagtactac caaaaggctg
     2281 aagaggcctg ggcccagatg gaggagtacc caactttctt gtacaaagtt ggcattataa
     2341 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     2401 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     2461 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2521 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     2581 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     2641 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     2701 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     2761 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     2821 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2881 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     2941 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     3001 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     3061 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     3121 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     3181 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     3241 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     3301 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     3361 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     3421 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     3481 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     3541 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     3601 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     3661 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     3721 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3781 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3841 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3901 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3961 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4021 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4081 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4141 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4201 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4261 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4321 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4381 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4441 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4501 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4561 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4621 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     4681 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     4741 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     4801 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     4861 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     4921 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgtt
//
