LOCUS       sequence-344653-        4227 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 4227)
  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 4227)
  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-CAMK2A
FEATURES             Location/Qualifiers
     source          1..4227
                     /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(1570..1669)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1686..1705)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1687..1705)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1710..1726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1710..1726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2157..2948
                     /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      3041..3629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3530..3549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3783..3800
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3959..3986)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4078..4164)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4213..4227,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4227,1..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 caccatggcc accatcacct gcacccgctt cacggaagag taccagctct
      181 tcgaggaatt gggcaaggga gccttctcgg tggtgcgaag gtgtgtgaag gtgctggctg
      241 gccaggagta tgctgccaag atcatcaaca caaagaagct gtcagccaga gaccatcaga
      301 agctggagcg tgaagcccgc atctgccgcc tgctgaagca ccccaacatc gtccgactac
      361 atgacagcat ctcagaggag ggacaccact acctgatctt cgacctggtc actggtgggg
      421 aactgtttga agatatcgtg gcccgggagt attacagtga ggcggatgcc agtcactgta
      481 tccagcagat cctggaggct gtgctgcact gccaccagat gggggtggtg caccgggacc
      541 tgaagcctga gaatctgttg ctggcctcca agctcaaggg tgccgcagtg aagctggcag
      601 actttggcct ggccatagag gtggaggggg agcagcaggc atggtttggg tttgcaggga
      661 ctcctggata tctctcccca gaagtgctgc ggaaggaccc gtacgggaag cctgtggacc
      721 tgtgggcttg tggggtcatc ctgtacatcc tgctggttgg gtaccccccg ttctgggatg
      781 aggaccagca ccgcctgtac cagcagatca aagccggcgc ctatgatttc ccatcgccgg
      841 aatgggacac tgtcaccccg gaagccaagg atctgatcaa taagatgctg accattaacc
      901 catccaaacg catcacagct gccgaagccc ttaagcaccc ctggatctcg caccgctcca
      961 ccgtggcatc ctgcatgcac agacaggaga ccgtggactg cctgaagaag ttcaatgcca
     1021 ggaggaaact gaagggagcc attctcacca cgatgctggc caccaggaac ttctccggag
     1081 ggaagagtgg gggaaacaag aagagcgatg gtgtgaagga atcctcagag agcaccaaca
     1141 ccaccatcga ggatgaagac accaaagtgc ggaaacagga aattataaaa gtgacagagc
     1201 agctgattga agccataagc aatggagatt ttgagtccta cacgaagatg tgcgaccctg
     1261 gcatgacagc cttcgaacct gaggccctgg ggaacctggt tgagggcctg gacttccatc
     1321 gattctattt tgaaaacctg tggtcccgga acagcaagcc cgtgcacacc accatcctga
     1381 atccccacat ccacctgatg ggcgacgagt cagcctgcat cgcctacatc cgcatcacgc
     1441 agtacctgga cgctggcggc atcccacgca ccgcccagtc ggaggagacc cgtgtctggc
     1501 accgccggga cggcaaatgg cagatcgtcc acttccacag atctggggcg ccctccgtcc
     1561 tgccccactt gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1621 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1681 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1741 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     1801 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     1861 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     1921 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1981 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     2041 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     2101 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     2161 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     2221 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     2281 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     2341 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     2401 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     2461 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     2521 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     2581 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     2641 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     2701 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     2761 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     2821 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     2881 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     2941 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3001 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3061 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3121 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3181 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3241 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3301 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3361 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3421 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3481 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3541 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3601 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3661 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3721 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3781 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3841 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3901 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     3961 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     4021 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     4081 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     4141 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     4201 catggatgtt ttcccagtca cgacgtt
//
