LOCUS       sequence-215967-        6252 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 6252)
  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 6252)
  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-JAK1
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(274..301)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(393..479)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     528..550
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     542..559
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     543..559
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    575..674
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4253..4272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4254..4272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4277..4293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4277..4293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4724..5515
                     /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      5608..6196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6097..6116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      241 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      301 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      361 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      421 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      481 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      541 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
      601 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
      661 caaaaaagtt ggcatgcagt atctaaatat aaaagaggac tgcaatgcca tggctttctg
      721 tgctaaaatg aggagctcca agaagactga ggtgaacctg gaggcccctg agccaggggt
      781 ggaagtgatc ttctatctgt cggacaggga gcccctccgg ctgggcagtg gagagtacac
      841 agcagaggaa ctgtgcatca gggctgcaca ggcatgccgt atctctcctc tttgtcacaa
      901 cctctttgcc ctgtatgacg agaacaccaa gctctggtat gctccaaatc gcaccatcac
      961 cgttgatgac aagatgtccc tccggctcca ctaccggatg aggttctatt tcaccaattg
     1021 gcatggaacc aacgacaatg agcagtcagt gtggcgtcat tctccaaaga agcagaaaaa
     1081 tggctacgag aaaaaaaaga ttccagatgc aacccctctc cttgatgcca gctcactgga
     1141 gtatctgttt gctcagggac agtatgattt ggtgaaatgc ctggctccta ttcgagaccc
     1201 caagaccgag caggatggac atgatattga gaacgagtgt ctagggatgg ctgtcctggc
     1261 catctcacac tatgccatga tgaagaagat gcagttgcca gaactgccca aggacatcag
     1321 ctacaagcga tatattccag aaacattgaa taagtccatc agacagagga accttctcac
     1381 caggatgcgg ataaataatg ttttcaagga tttcctaaag gaatttaaca acaagaccat
     1441 ttgtgacagc agcgtgtcca cgcatgacct gaaggtgaaa tacttggcta ccttggaaac
     1501 tttgacaaaa cattacggtg ctgaaatatt tgagacttcc atgttactga tttcatcaga
     1561 aaatgagatg aattggtttc attcgaatga cggtggaaac gttctctact acgaagtgat
     1621 ggtgactggg aatcttggaa tccagtggag gcataaacca aatgttgttt ctgttgaaaa
     1681 ggaaaaaaat aaactgaagc ggaaaaaact ggaaaataaa cacaagaagg atgaggagaa
     1741 aaacaagatc cgggaagagt ggaacaattt ttcttacttc cctgaaatca ctcacattgt
     1801 aataaaggag tctgtggtca gcattaacaa gcaggacaac aagaaaatgg aactgaagct
     1861 ctcttcccac gaggaggcct tgtcctttgt gtccctggta gatggctact tccggctcac
     1921 agcagatgcc catcattacc tctgcaccga cgtggccccc ccgttgatcg tccacaacat
     1981 acagaatggc tgtcatggtc caatctgtac agaatacgcc atcaataaat tgcggcaaga
     2041 aggaagcgag gaggggatgt acgtgctgag gtggagctgc accgactttg acaacatcct
     2101 catgaccgtc acctgctttg agaagtctga gcaggtgcag ggtgcccaga agcagttcaa
     2161 gaactttcag atcgaggtgc agaagggccg ctacagtctg cacggttcgg accgcagctt
     2221 ccccagcttg ggagacctca tgagccacct caagaagcag atcctgcgca cggataacat
     2281 cagcttcatg ctaaaacgct gctgccagcc caagccccga gaaatctcca acctgctggt
     2341 ggctactaag aaagcccagg agtggcagcc cgtctacccc atgagccagc tgagtttcga
     2401 tcggatcctc aagaaggatc tggtgcaggg cgagcacctt gggagaggca cgagaacaca
     2461 catctattct gggaccctga tggattacaa ggatgacgaa ggaacttctg aagagaagaa
     2521 gataaaagtg atcctcaaag tcttagaccc cagccacagg gatatttccc tggccttctt
     2581 cgaggcagcc agcatgatga gacaggtctc ccacaaacac atcgtgtacc tctatggcgt
     2641 ctgtgtccac gacgtggaga atatcatggt ggaagagttt gtggaagggg gtcctctgga
     2701 tctcttcatg caccggaaaa gcgatgtcct taccacacca tggaaattca aagttgccaa
     2761 acagctggcc agtgccctga gctacttgga ggataaagac ctggtccatg gaaatgtgtg
     2821 tactaaaaac ctcctcctgg cccgtgaggg catcgacagt gagtgtggcc cattcatcaa
     2881 gctcagtgac cccggcatcc ccattacggt gctgtctagg caagaatgca ttgaacgaat
     2941 cccatggatt gctcctgagt gtgttgagga ctccaagaac ctgagtgtgg ctgctgacaa
     3001 gtggagcttt ggaaccacgc tctgggaaat ctgctacaat ggcgagatcc ccttgaaaga
     3061 caagacgctg attgagaaag agagattcta tgaaagccgg tgcaggccag tgacaccatc
     3121 atgtaaggag ctggctgacc tcatgacccg ctgcatgaac tatgacccca atcagaggcc
     3181 tttcttccga gccatcatga gagacattaa taagcttgaa gagcagaatc cagatattgt
     3241 ttcagaaaaa aaaccagcaa ctgaagtgga ccccacacat tttgaaaagc gcttcctaaa
     3301 gaggatccgt gacttgggag agggccactt tgggaaggtt gagctctgca ggtatgaccc
     3361 cgaaggggac aatacagggg agcaggtggc tgttaaatct ctgaagcctg agagtggagg
     3421 taaccacata gctgatctga aaaaggaaat cgagatctta aggaacctct atcatgagaa
     3481 cattgtgaag tacaaaggaa tctgcacaga agacggagga aatggtatta agctcatcat
     3541 ggaatttctg ccttcgggaa gccttaagga atatcttcca aagaataaga acaaaataaa
     3601 cctcaaacag cagctaaaat atgccgttca gatttgtaag gggatggact atttgggttc
     3661 tcggcaatac gttcaccggg acttggcagc aagaaatgtc cttgttgaga gtgaacacca
     3721 agtgaaaatt ggagacttcg gtttaaccaa agcaattgaa accgataagg agtattacac
     3781 cgtcaaggat gaccgggaca gccctgtgtt ttggtatgct ccagaatgtt taatgcaatc
     3841 taaattttat attgcctctg acgtctggtc ttttggagtc actctgcatg agctgctgac
     3901 ttactgtgat tcagattcta gtcccatggc tttgttcctg aaaatgatag gcccaaccca
     3961 tggccagatg acagtcacaa gacttgtgaa tacgttaaaa gaaggaaaac gcctgccgtg
     4021 cccacctaac tgtccagatg aggtttatca acttatgagg aaatgctggg aattccaacc
     4081 atccaatcgg acaagctttc agaaccttat tgaaggattt gaagcacttt taaaattgcc
     4141 aactttcttg tccaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     4201 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     4261 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     4321 ctgatgttac attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga
     4381 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     4441 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     4501 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     4561 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     4621 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     4681 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
     4741 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     4801 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     4861 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     4921 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     4981 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     5041 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     5101 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     5161 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     5221 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     5281 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     5341 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     5401 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     5461 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     5521 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6241 gccttttgct ca
//
