LOCUS       sequence-142383-        6351 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 6351)
  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 6351)
  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-TYK2
FEATURES             Location/Qualifiers
     source          1..6351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4111..4210)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4227..4246)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4228..4246)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4251..4267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4251..4267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4698..5489
                     /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      5582..6170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6071..6090
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6324..6341
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcac catgcctctg cgccactggg ggatggccag gggcagtaag cccgttgggg
      601 atggagccca gcccatggct gccatgggag gcctgaaggt gcttctgcac tgggctggtc
      661 caggcggcgg ggagccctgg gtcactttca gtgagtcatc gctgacagct gaggaagtct
      721 gcatccacat tgcacataaa gttggtatca ctcctccttg cttcaatctc tttgccctct
      781 tcgatgctca ggcccaagtc tggttgcccc caaaccacat cctagagatc cccagagatg
      841 caagcctgat gctatatttc cgcataaggt tttatttccg gaactggcat ggcatgaatc
      901 ctcgggaacc ggctgtgtac cgttgtgggc ccccaggaac cgaggcatcc tcagatcaga
      961 cagcacaggg gatgcaactc ctggacccag cctcatttga gtacctcttt gagcagggca
     1021 agcatgagtt tgtgaatgac gtggcatcac tgtgggagct gtcgaccgag gaggagatcc
     1081 accactttaa gaatgagagc ctgggcatgg cctttctgca cctctgtcac ctcgctctcc
     1141 gccatggcat ccccctggag gaggtggcca agaagaccag cttcaaggac tgcatcccgc
     1201 gctccttccg ccggcatatc cggcagcaca gcgccctgac ccggctgcgc cttcggaacg
     1261 tcttccgcag gttcctgcgg gacttccagc cgggccgact ctcccagcag atggtcatgg
     1321 tcaaatacct agccacactc gagcggctgg caccccgctt cggcacagag cgtgtgcccg
     1381 tgtgccacct gaggctgctg gcccaggccg agggggagcc ctgctacatc cgggacagtg
     1441 gggtggcccc tacagaccct ggccctgagt ctgctgctgg gcccccaacc cacgaggtgc
     1501 tggtgacagg cactggtggc atccagtggt ggccagtaga ggaggaggtg aacaaggagg
     1561 agggttctag tggcagcagt ggcaggaacc cccaagccag cctgtttggg aagaaggcca
     1621 aggctcacaa ggcagtcggc cagccggcag acaggccgcg ggagccactg tgggcctact
     1681 tctgtgactt ccgggacatc acccacgtgg tgctgaaaga gcactgtgtc agcatccacc
     1741 ggcaggacaa caagtgcctg gagctgagct tgccttcccg ggctgcggcg ctgtccttcg
     1801 tgtcgctggt ggacggctat ttccgcctga cggccgactc cagccactac ctgtgccacg
     1861 aggtggctcc cccacggctg gtgatgagca tccgggatgg gatccacgga cccctgctgg
     1921 agccatttgt gcaggccaag ctgcggcccg aggacggcct gtacctcatt cactggagca
     1981 ccagccaccc ctaccgcctg atcctcacag tggcccagcg tagccaggca ccagacggca
     2041 tgcagagctt gcggctccga aagttcccca ttgagcagca ggacggggcc ttcgtgctgg
     2101 agggctgggg ccggtccttc cccagcgttc gggaacttgg ggctgccttg cagggctgct
     2161 tgctgagggc cggggatgac tgcttctctc tgcgtcgctg ttgcctgccc caaccaggag
     2221 aaacctccaa tctcatcatc atgcgggggg ctcgggccag ccccaggaca ctcaacctca
     2281 gccagctcag cttccaccgg gttgaccaga aggagatcac ccagctgtcc cacttgggcc
     2341 agggcacaag gaccaacgtg tatgagggcc gcctgcgagt ggagggcagc ggggaccctg
     2401 aggagggcaa gatggatgac gaggaccccc tcgtgcctgg cagggaccgt gggcaggagc
     2461 tacgagtggt gctcaaagtg ctggacccta gtcaccatga catcgccctg gccttctacg
     2521 agacagccag cctcatgagc caggtctccc acacgcacct ggccttcgtg catggcgtct
     2581 gtgtgcgcgg ccctgaaaat atcatggtga cagagtacgt ggagcacgga cccctggatg
     2641 tgtggctgcg gagggagcgg ggccatgtgc ccatggcttg gaagatggtg gtggcccagc
     2701 agctggccag cgccctcagc tacctggaga acaagaacct ggttcatggt aatgtgtgtg
     2761 gccggaacat cctgctggcc cggctggggt tggcagaggg caccagcccc ttcatcaagc
     2821 tgagtgatcc tggcgtgggc ctgggcgccc tctccaggga ggagcgggtg gagaggatcc
     2881 cctggctggc ccccgaatgc ctaccaggtg gggccaacag cctaagcacc gccatggaca
     2941 agtgggggtt tggcgccacc ctcctggaga tctgctttga cggagaggcc cctctgcaga
     3001 gccgcagtcc ctccgagaag gagcatttct accagaggca gcaccggctg cccgagccct
     3061 cctgcccaca gctggccaca ctcaccagcc agtgtctgac ctatgagcca acccagaggc
     3121 catcattccg caccatcctg cgtgacctca cccggctgca gccccacaat cttgctgacg
     3181 tcttgactgt gaacccggac tcaccggcgt cggaccctac ggttttccac aagcgctatt
     3241 tgaaaaagat ccgagatctg ggcgagggtc acttcggcaa ggtcagcttg tactgctacg
     3301 atccgaccaa cgacggcact ggcgagatgg tggcggtgaa agccctcaag gcagactgcg
     3361 gcccccagca ccgctcgggc tggaagcagg agattgacat tctgcgcacg ctctaccacg
     3421 agcacatcat caagtacaag ggctgctgcg aggaccaagg cgagaagtcg ctgcagctgg
     3481 tcatggagta cgtgcccctg ggcagcctcc gagactacct gccccggcac agcatcgggc
     3541 tggcccagct gctgctcttc gcccagcaga tctgcgaggg catggcctat ctgcactcgc
     3601 agcactacat ccaccgagac ctagccgcgc gcaacgtgct gctggacaac gacaggctgg
     3661 tcaagatcgg ggactttggc ctagccaagg ccgtgcccga aggccacgag tactaccgcg
     3721 tgcgcgagga tggggacagc cccgtgttct ggtatgcccc agagtgcctg aaggagtata
     3781 agttctacta tgcgtcagat gtctggtcct tcggggtgac cctgtatgag ctgctgacgc
     3841 actgtgactc cagccagagc ccccccacga aattccttga gctcataggc attgctcagg
     3901 gtcagatgac agttctgaga ctcactgagt tgctggaacg aggggagagg ctgccacggc
     3961 ccgacaaatg tccctgtgag gtctatcatc tcatgaagaa ctgctgggag acagaggcgt
     4021 cctttcgccc aaccttcgag aacctcatac ccattctgaa gacagtccat gagaagtacc
     4081 aaggccaggc cccttcagtg ttcagcgtgt gcccaacttt cttgtacaaa gttggcatta
     4141 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     4201 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     4261 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     4321 atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
     4381 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     4441 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     4501 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     4561 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     4621 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     4681 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     4741 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     4801 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     4861 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     4921 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     4981 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     5041 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     5101 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     5161 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     5221 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     5281 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     5341 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     5401 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     5461 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     5521 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5701 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5761 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5821 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5881 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5941 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6001 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6061 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
