LOCUS       sequence-349164-        4860 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 4860)
  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 4860)
  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-LATS1
FEATURES             Location/Qualifiers
     source          1..4860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2171..2270)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2287..2306)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2288..2306)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2311..2327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2311..2327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2758..3549
                     /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      3642..4230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4131..4150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4384..4401
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4560..4587)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4679..4765)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4814..4836
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4828..4845
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4829..4845
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagttggca tgaagaggag tgaaaagcca
      121 gaaggatata gacaaatgag gcctaagacc tttcctgcca gtaactatac tgtcagtagc
      181 cggcaaatgt tacaagaaat tcgggaatcc cttaggaatt tatctaaacc atctgatgct
      241 gctaaggctg agcataacat gagtaaaatg tcaaccgaag atcctcgaca agtcagaaat
      301 ccacccaaat ttgggacgca tcataaagcc ttgcaggaaa ttcgaaactc tctgcttcca
      361 tttgcaaatg aaacaaattc ttctcggagt acttcagaag ttaatccaca aatgcttcaa
      421 gacttgcaag ctgctggatt tgatgaggat atggttatac aagctcttca gaaaactaac
      481 aacagaagta tagaagcagc aattgaattc attagtaaaa tgagttacca agatcctcga
      541 cgagagcaga tggctgcagc agctgccaga cctattaatg ccagcatgaa accagggaat
      601 gtgcagcaat cagttaaccg caaacagagc tggaaaggtt ctaaagaatc cttagttcct
      661 cagaggcatg gcccgccact aggagaaagt gtggcctatc attctgagag tcccaactca
      721 cagacagatg taggaagacc tttgtctgga tctggtatat cagcatttgt tcaagctcac
      781 cctagcaacg gacagagagt gaacccccca ccaccacctc aagtaaggag tgttactcct
      841 ccaccacctc caagaggcca gactccccct ccaagaggta caactccacc tcccccttca
      901 tgggaaccaa actctcaaac aaagcgctat tctggaaaca tggaatacgt aatctcccga
      961 atctctcctg tcccacctgg ggcatggcaa gagggctatc ctccaccacc tctcaacact
     1021 tcccccatga atcctcctaa tcaaggacag agaggcatta gttctgttcc tgttggcaga
     1081 caaccaatca tcatgcagag ttctagcaaa tttaactttc catcagggag acctggaatg
     1141 cagaatggta ctggacaaac tgatttcatg atacaccaaa atgttgtccc tgctggcact
     1201 gtgaatcggc agccaccacc tccatatcct ctgacagcag ctaatggaca aagcccttct
     1261 gctttacaaa cagggggatc tgctgctcct tcgtcatata caaatggaag tattcctcag
     1321 tctatgatgg tgccaaacag aaatagtcat aacatggaac tatataacat tagtgtacct
     1381 ggactgcaaa caaattggcc tcagtcatct tctgctccag cccagtcatc cccgagcagt
     1441 gggcatgaaa tccctacatg gcaacctaac ataccagtga ggtcaaattc ttttaataac
     1501 ccattaggaa atagagcaag tcactctgct aattctcagc cttccgctac aacagtcact
     1561 gcaattacac cagctcctat tcaacagcct gtgaaaagta tgcgtgtatt aaaaccagag
     1621 ctacagactg ctttagcacc tacacaccct tcttggatac cacagccaat tcaaactgtt
     1681 caacccagtc cttttcctga gggaaccgct tcaaatgtga ctgtgatgcc acctgttgct
     1741 gaagctccaa actatcaagg accaccacca ccctacccaa aacatctgct gcaccaaaac
     1801 ccatctgttc ctccatacga gtcaatcagt aagcctagca aagaggatca gccaagcttg
     1861 cccaaggaag atgagagtga aaagagttat gaaaatgttg atagtgggga taaagaaaag
     1921 aaacagatta caacttcacc tattactgtt aggaaaaaca agaaagatga agagcgaagg
     1981 gaatctcgta ttcaaagtta ttctcctcaa gcatttaaat tctttatgga gcaacatgta
     2041 gaaaatgtac tcaaatctca tcagcagcgt ctacatcgta aaaaacaatt agagaatgaa
     2101 atgatgcggg taaaaccttt taaaatgtcc atttttatac ttaatcatct gtttgcttgg
     2161 tgtttatttt acccaacttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     2221 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     2281 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2341 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgcctcctc
     2401 tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
     2461 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
     2521 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     2581 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
     2641 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     2701 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
     2761 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
     2821 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
     2881 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
     2941 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
     3001 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
     3061 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
     3121 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
     3181 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
     3241 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
     3301 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     3361 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     3421 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     3481 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     3541 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     3601 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3661 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3721 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3781 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3841 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3901 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3961 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4021 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4081 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4141 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4201 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4261 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4321 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4381 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4441 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4501 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     4561 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     4621 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     4681 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     4741 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     4801 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
