LOCUS       sequence-251772-        4695 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 4695)
  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 4695)
  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-SYK
FEATURES             Location/Qualifiers
     source          1..4695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     558..575
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(734..761)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(853..939)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     988..1010
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1002..1019
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1003..1019
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1035..1134
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3040..3139)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3156..3175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3157..3175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3180..3196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3180..3196)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3627..4418
                     /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      join(4511..4695,1..404)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
       61 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      121 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      181 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      241 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      301 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      361 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      421 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      481 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
      541 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
      601 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      661 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
      721 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
      781 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
      841 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
      901 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
      961 tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc
     1021 ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt tcgttgcaac
     1081 aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt ggcaccatgg
     1141 ccagcagcgg catggctgac agcgccaacc acctgccctt ctttttcggc aacatcaccc
     1201 gggaggaggc agaagattac ctggtccagg ggggcatgag tgatgggctt tatttgctgc
     1261 gccagagccg caactacctg ggtggcttcg ccctgtccgt ggcccacggg aggaaggcac
     1321 accactacac catcgagcgg gagctgaatg gcacctacgc catcgccggt ggcaggaccc
     1381 atgccagccc cgccgacctc tgccactacc actcccagga gtctgatggc ctggtctgcc
     1441 tcctcaagaa gcccttcaac cggccccaag gggtgcagcc caagactggg ccctttgagg
     1501 atttgaagga aaacctcatc agggaatatg tgaagcagac atggaacctg cagggtcagg
     1561 ctctggagca ggccatcatc agtcagaagc ctcagctgga gaagctgatc gctaccacag
     1621 cccatgaaaa aatgccttgg ttccatggaa aaatctctcg ggaagaatct gagcaaattg
     1681 tcctgatagg atcaaagaca aatggaaagt tcctgatccg agccagagac aacaacggct
     1741 cctacgccct gtgcctgctg cacgaaggga aggtgctgca ctatcgcatc gacaaagaca
     1801 agacagggaa gctctccatc cccgagggaa agaagttcga cacgctctgg cagctagtcg
     1861 agcattattc ttataaagca gatggtttgt taagagttct tactgtccca tgtcaaaaaa
     1921 tcggcacaca gggaaatgtt aattttggag gccgtccaca acttccaggt tcccatcctg
     1981 cgacttggtc agcgggtgga ataatctcaa gaatcaaatc atactccttc ccaaagcctg
     2041 gccacagaaa gtcctcccct gcccaaggga accggcaaga gagtactgtg tcattcaatc
     2101 cgtatgagcc agaacttgca ccctgggctg cagacaaagg cccccagaga gaagccctac
     2161 ccatggacac agaggtgtac gagagcccct acgcggaccc tgaggagatc aggcccaagg
     2221 aggtttacct ggaccgaaag ctgctgacgc tggaagacaa agaactgggc tctggtaatt
     2281 ttggaactgt gaaaaagggc tactaccaaa tgaaaaaagt tgtgaaaacc gtggctgtga
     2341 aaatactgaa aaacgaggcc aatgaccccg ctcttaaaga tgagttatta gcagaagcaa
     2401 atgtcatgca gcagctggac aacccgtaca tcgtgcgcat gatcgggata tgcgaggccg
     2461 agtcctggat gctagttatg gagatggcag aacttggtcc cctcaataag tatttgcagc
     2521 agaacagaca tgtcaaggat aagaacatca tagaactggt tcatcaggtt tccatgggca
     2581 tgaagtactt ggaggagagc aattttgtgc acagagatct ggctgcaaga aatgtgttgc
     2641 tagttaccca acattatgcc aagatcagtg atttcggact ctccaaagca ctgcgtgctg
     2701 atgaaaacta ctacaaggcc cagacccatg gaaagtggcc tgtcaagtgg tacgctccgg
     2761 aatgcatcaa ctactacaag ttctccagca aaagcgatgt ctggagcttt ggagtgttga
     2821 tgtgggaagc attctcctat gggcagaagc catatcgagg gatgaaagga agtgaagtca
     2881 ccgctatgtt agagaaagga gagcggatgg ggtgccctgc agggtgtcca agagagatgt
     2941 acgatctcat gaatctgtgc tggacatacg atgtggaaaa caggcccgga ttcgcagcag
     3001 tggaactgcg gctgcgcaat tactactatg acgtggtgaa cccaactttc ttgtacaaag
     3061 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     3121 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     3181 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     3241 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     3301 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3361 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3421 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3481 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3541 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     3601 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     3661 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     3721 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     3781 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     3841 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     3901 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     3961 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     4021 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     4081 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4141 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4201 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     4261 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     4321 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     4381 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     4441 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4621 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4681 gccaccactt caaga
//
