LOCUS       sequence-271786-        4044 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 4044)
  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 4044)
  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-PRKAR2B
FEATURES             Location/Qualifiers
     source          1..4044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..88
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     72..88
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     93..112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        93..111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    129..228
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1483..1582)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1598..1615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1598..1614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1678..1764
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1856..1883
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2042..2059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2213..2801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2293..2312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2894..3685)
                     /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"
ORIGIN
        1 ggaggcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg
       61 gccagagctg ccaggaaaca gctatgacca tgtaatacga ctcactatag gggatatcag
      121 ctggatggca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg
      181 ataagcaatg ctttcttata atgccaactt tgtacaagaa agttgggcat gcagtgggtt
      241 caacaatatc catgttcgtt ccaaacaggg caactaactg ttcttcatag gtagcgatgt
      301 tccttttcat aatttccatg caaggtccca gaagcctttc aaatgcttgc acatccattg
      361 ctaaacattt gacagtccca atggcgtggg cagaagctgc tcgaggtttg ttagttacca
      421 gggcaagctc tccaaagtac tgtccccgcg agcatcgagc gatttctact gcaccattct
      481 cttccacttc tgatttaccc tttcttttca tagtaatttt cacttctcca gattctacaa
      541 tgaaaaaaga atcagccgaa tctccctgag caatgatttg ttctccatcg ttgtatactt
      601 tggtgcctat cacatctact actttcaggc gttcagaaaa ctccaaagat ttaaggaatg
      661 gcagtgactc aataaagctt tcatacattt ttctcttttt ggcattgttt ttcacaatta
      721 ttctcctgaa ggttaccctg tccaaacccc acagagcacc aggagaggta gcagtgattg
      781 tagctgctct gggtgtattg tacattaagg ccagttcgcc gaaactccca cgattatcat
      841 agttaccaac acatcttcca acaccatcac atttcacata aatatcaaat gtgcctctat
      901 caattacata aaagttgtca ccatcgtcac cttgatcaat tacatgctcc ccatctttga
      961 ccaatttttc aaacatggca tctaatactt gagacatctg ctccggatcc agattcttaa
     1021 acagcaggat gtctttgcaa gcctcttgca acctatttct ttgatcatca gtttttggat
     1081 gtataatcct ggactctgca tcatcttctt cttcatcagg attataagct tctgcacata
     1141 ctgaggcacg ccttgtgaat cggtttatta ctggagcatt gaacgcccct gcgtccgcgg
     1201 gcgccgcctc ctcctcctcc ccgtcctcgg agtcggactg catgggctcc tcggcgaagt
     1261 tgaccccctt gctgggggtg ccgcccccgg cagcggcgcc caggtccccc caggtcctgc
     1321 cctcatggca gaagcgcgcg gtgcctttgc gctcgttctc ctgctgcagg cgggtgaagt
     1381 gctgcagagc gaactccagc aggtccgcgg gctggtgcct cagcacctcc accgtgaagc
     1441 cctgcagcag ctccgtcagt cccgccggga tctcgatgct catgccaact tttttgtaca
     1501 aagttggcat tataaaaaag cattgctcat caatttgttg caacgaacag gtcactatca
     1561 gtcaaaataa aatcattatt tggggcccga gcttaagact ggccgtcgtt ttacaacgtc
     1621 gtgactggga aaacatccat gctagcgtta acgcgagagt agggaactgc caggcatcaa
     1681 ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1741 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg
     1801 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag
     1861 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg
     1921 tattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2821 ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat tttggtcatg
     2881 ggtggctcga gggttatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa
     2941 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
     3001 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
     3061 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
     3121 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
     3181 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
     3241 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
     3301 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
     3361 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
     3421 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
     3481 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     3541 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     3601 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     3661 cttcggcgat caccgcttcc ctcataatgt ttaactttgt tttagggcga ctgccctgct
     3721 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     3781 gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa
     3841 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     3901 gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg
     3961 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggtagcagc gaagtcgagg
     4021 catttctgtc ctggctggtc taga
//
