LOCUS       sequence-335008-        6165 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 6165)
  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 6165)
  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-JAK3
FEATURES             Location/Qualifiers
     source          1..6165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(93..120)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(212..298)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     347..369
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     361..378
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     362..378
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    394..493
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3869..3968)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3985..4004)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3986..4004)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4009..4025)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4009..4025)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4456..5247
                     /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      5340..5928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5829..5848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6082..6099
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
       61 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
      121 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
      181 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
      241 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
      301 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
      361 tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata
      421 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac
      481 aaaaaagttg gcaccatggc acctccaagt gaagagacgc ccctgatccc tcagcgttca
      541 tgcagcctct tgtccacgga ggctggtgcc ctgcatgtgc tgctgcccgc tcggggcccc
      601 gggccccccc agcgcctatc tttctccttt ggggaccact tggctgagga cctgtgcgtg
      661 caggctgcca aggccagcgg catcctgcct gtgtaccact ccctctttgc tctggccacg
      721 gaggacctgt cctgctggtt ccccccgagc cacatcttct ccgtggagga tgccagcacc
      781 caagtcctgc tgtacaggat tcgcttttac ttccccaatt ggtttgggct ggagaagtgc
      841 caccgcttcg ggctacgcaa ggatttggcc agtgctatcc ttgacctgcc agtcctggag
      901 cacctctttg cccagcaccg cagtgacctg gtgagtgggc gcctccccgt gggcctcagt
      961 ctcaaggagc agggtgagtg tctcagcctg gccgtgttgg acctggcccg gatggcgcga
     1021 gagcaggccc agcggccggg agagctgctg aagactgtca gctacaaggc ctgcctaccc
     1081 ccaagcctgc gcgacctgat ccagggcctg agcttcgtga cgcggaggcg tattcggagg
     1141 acggtgcgca gagccctgcg ccgcgtggcc gcctgccagg cagaccggca ctcgctcatg
     1201 gccaagtaca tcatggacct ggagcggctg gatccagccg gggccgccga gaccttccac
     1261 gtgggcctcc ctggggccct tggtggccac gacgggctgg ggctgctccg cgtggctggt
     1321 gacggcggca tcgcctggac ccagggagaa caggaggtcc tccagccctt ctgcgacttt
     1381 ccagaaatcg tagacattag catcaagcag gccccgcgcg ttggcccggc cggagagcac
     1441 cgcctggtca ctgttaccag gacagacaac cagattttag aggccgagtt cccagggctg
     1501 cccgaggctc tgtcgttcgt ggcgctcgtg gacggctact tccggctgac cacggactcc
     1561 cagcacttct tctgcaagga ggtggcaccg ccgaggctgc tggaggaagt ggccgagcag
     1621 tgccacggcc ccatcactct ggactttgcc atcaacaagc tcaagactgg gggctcacgt
     1681 cctggctcct atgttctccg ccgcagcccc caggactttg acagcttcct cctcactgtc
     1741 tgtgtccaga acccccttgg tcctgattat aagggctgcc tcatccggcg cagccccaca
     1801 ggaaccttcc ttctggttgg cctcagccga ccccacagca gtcttcgaga gctcctggca
     1861 acctgctggg atggggggct gcacgtagat ggggtggcag tgaccctcac ttcctgctgt
     1921 atccccagac ccaaagaaaa gtccaacctg atcgtggtcc agagaggtca cagcccaccc
     1981 acatcatcct tggttcagcc ccaatcccaa taccagctga gtcagatgac atttcacaag
     2041 atccctgctg acagcctgga gtggcatgag aacctgggcc atgggtcctt caccaagatt
     2101 taccggggct gtcgccatga ggtggtggat ggggaggccc gaaagacaga ggtgctgctg
     2161 aaggtcatgg atgccaagca caagaactgc atggagtcat tcctggaagc agcgagcttg
     2221 atgagccaag tgtcgtaccg gcatctcgtg ctgctccacg gcgtgtgcat ggctggagac
     2281 agcaccatgg tgcaggaatt tgtacacctg ggggccatag acatgtatct gcgaaaacgt
     2341 ggccacctgg tgccagccag ctggaagctg caggtggtca aacagctggc ctacgccctc
     2401 aactatctgg aggacaaagg cctgccccat ggcaatgtct ctgcccggaa ggtgctcctg
     2461 gctcgggagg gggctgatgg gagcccgccc ttcatcaagc tgagtgaccc tggggtcagc
     2521 cccgctgtgt taagcctgga gatgctcacc gacaggatcc cctgggtggc ccccgagtgt
     2581 ctccgggagg cgcagacact tagcttggaa gctgacaagt ggggcttcgg cgccacggtc
     2641 tgggaagtgt ttagtggcgt caccatgccc atcagtgccc tggatcctgc taagaaactc
     2701 caattttatg aggaccggca gcagctgccg gcccccaagt ggacagagct ggccctgctg
     2761 attcaacagt gcatggccta tgagccggtc cagaggccct ccttccgagc cgtcattcgt
     2821 gacctcaata gcctcatctc ttcagactat gagctcctct cagaccccac acctggtgcc
     2881 ctggcacctc gtgatgggct gtggaatggt gcccagctct atgcctgcca agaccccacg
     2941 atcttcgagg agagacacct caagtacatc tcacagctgg gcaagggcaa ctttggcagc
     3001 gtggagctgt gccgctatga cccgctaggc gacaatacag gtgccctggt ggccgtgaaa
     3061 cagctgcagc acagcgggcc agaccagcag agggactttc agcgggagat tcagatcctc
     3121 aaagcactgc acagtgattt cattgtcaag tatcgtggtg tcagctatgg cccgggccgc
     3181 cagagcctgc ggctggtcat ggagtacctg cccagcggct gcttgcgcga cttcctgcag
     3241 cggcaccgcg cgcgcctcga tgccagccgc ctccttctct attcctcgca gatctgcaag
     3301 ggcatggagt acctgggctc ccgccgctgc gtgcaccgcg acctggccgc ccgaaacatc
     3361 ctcgtggaga gcgaggcaca cgtcaagatc gctgacttcg gcctagctaa gctgctgccg
     3421 cttgacaaag actactacgt ggtccgcgag ccaggccaga gccccatttt ctggtatgcc
     3481 cccgaatccc tctcggacaa catcttctct cgccagtcag acgtctggag cttcggggtc
     3541 gtcctgtacg agctcttcac ctactgcgac aaaagctgca gcccctcggc cgagttcctg
     3601 cggatgatgg gatgtgagcg ggatgtcccc gccctctgcc gcctcttgga actgctggag
     3661 gagggccaga ggctgccggc gcctcctgcc tgccctgctg aggttcacga gctcatgaag
     3721 ctgtgctggg cccctagccc acaggaccgg ccatcattca gcgccctggg cccccagctg
     3781 gacatgctgt ggagcggaag ccgggggtgt gagactcatg ccttcactgc tcacccagag
     3841 ggcaaacacc actccctgtc cttttcatag ccaactttct tgtacaaagt tggcattata
     3901 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     3961 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     4021 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     4081 atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc
     4141 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa
     4201 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     4261 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     4321 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     4381 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     4441 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     4501 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     4561 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     4621 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     4681 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     4741 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     4801 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     4861 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     4921 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     4981 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     5041 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     5101 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     5161 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     5221 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac
     5281 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5461 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5941 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6001 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6061 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6121 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagc
//
