LOCUS       sequence-343699-        4647 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 4647)
  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 4647)
  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-ZAP70
FEATURES             Location/Qualifiers
     source          1..4647
                     /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(1958..2057)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2074..2093)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2075..2093)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2098..2114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2098..2114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2545..3336
                     /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      3429..4017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3918..3937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4171..4188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4347..4374)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4466..4552)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4601..4623
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4615..4632
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4616..4632
                     /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 ccatgccaga ccccgcggcg
      121 cacctgccct tcttctacgg cagcatctcg cgtgccgagg ccgaggagca cctgaagctg
      181 gcgggcatgg cggacgggct cttcctgctg cgccagtgcc tgcgctcgct gggcggctat
      241 gtgctgtcgc tcgtgcacga tgtgcgcttc caccactttc ccatcgagcg ccagctcaac
      301 ggcacctacg ccattgccgg cggcaaagcg cactgtggac cggcagagct ctgcgagttc
      361 tactcgcgcg accccgacgg gctgccctgc aacctgcgca agccgtgcaa ccggccgtcg
      421 ggcctcgagc cgcagccggg ggtcttcgac tgcctgcgag acgccatggt gcgtgactac
      481 gtgcgccaga cgtggaagct ggagggcgag gccctggagc aggccatcat cagccaggcc
      541 ccgcaggtgg agaagctcat tgctacgacg gcccacgagc ggatgccctg gtaccacagc
      601 agcctgacgc gtgaggaggc cgagcgcaaa ctttactctg gggcgcagac cgacggcaag
      661 ttcctgctga ggccgcggaa ggagcagggc acatacgccc tgtccctcat ctatgggaag
      721 acggtgtacc actacctcat cagccaagac aaggcgggca agtactgcat tcccgagggc
      781 accaagtttg acacgctctg gcagctggtg gagtatctga agctgaaggc ggacgggctc
      841 atctactgcc tgaaggaggc ctgccccaac agcagtgcca gcaacgcctc aggggctgct
      901 gctcccacac tcccagccca cccatccacg ttgactcatc ctcagagacg aatcgacacc
      961 ctcaactcag atggatacac ccctgagcca gcacgcataa cgtccccaga caaaccgcgg
     1021 ccgatgccca tggacacgag cgtgtatgag agcccctaca gcgacccaga ggagctcaag
     1081 gacaagaagc tcttcctgaa gcgcgataac ctcctcatag ctgacattga acttggctgc
     1141 ggcaactttg gctcagtgcg ccagggcgtg taccgcatgc gcaagaagca gatcgacgtg
     1201 gccatcaagg tgctgaagca gggcacggag aaggcagaca cggaagagat gatgcgcgag
     1261 gcgcagatca tgcaccagct ggacaacccc tacatcgtgc ggctcattgg cgtctgccag
     1321 gccgaggccc tcatgctggt catggagatg gctgggggcg ggccgctgca caagttcctg
     1381 gtcggcaaga gggaggagat ccctgtgagc aatgtggccg agctgctgca ccaggtgtcc
     1441 atggggatga agtacctgga ggagaagaac tttgtgcacc gtgacctggc ggcccgcaac
     1501 gtcctgctgg ttaaccggca ctacgccaag atcagcgact ttggcctctc caaagcactg
     1561 ggtgccgacg acagctacta cactgcccgc tcagcaggga agtggccgct caagtggtac
     1621 gcacccgaat gcatcaactt ccgcaagttc tccagccgca gcgatgtctg gagctatggg
     1681 gtcaccatgt gggaggcctt gtcctacggc cagaagccct acaagaagat gaaagggccg
     1741 gaggtcatgg ccttcatcga gcagggcaag cggatggagt gcccaccaga gtgtccaccc
     1801 gaactgtacg cactcatgag tgactgctgg atctacaagt gggaggatcg ccccgacttc
     1861 ctgaccgtgg agcagcgcat gcgagcctgt tactacagcc tggccagcaa ggtggaaggg
     1921 cccccaggca gcacacagaa ggctgaggct gcctgtgccc caactttctt gtacaaagtt
     1981 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2041 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     2101 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     2161 gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg
     2221 ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag
     2281 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
     2341 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     2401 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
     2461 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
     2521 cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa
     2581 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
     2641 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
     2701 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
     2761 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
     2821 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
     2881 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
     2941 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
     3001 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
     3061 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     3121 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     3181 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     3241 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     3301 gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa
     3361 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3541 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4141 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4201 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4261 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     4321 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     4381 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     4441 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     4501 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     4561 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     4621 acgacggcca gtcttaagct cgggccc
//
