LOCUS       sequence-161647-        5094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing BRAF, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5094)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 5094)
  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: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_BRAF_p.G469V
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      179..188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(189..288)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(304..321)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(304..320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(313..335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      384..470
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      562..589
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(748..765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(919..1507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(999..1018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1600..2391)
                     /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"
     primer_bind     2822..2838
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2822..2838
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2843..2862
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2843..2861
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2879..2978
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ctctatatgt tcctgtgtca acttaatcat ttgtttgata ttccacacct cctccggaat
       61 ggcagggtcc gcagccgaag aggccgcggc gccggcgccg gcgccggcct cgggctccat
      121 gtccccgttg aacagagcct ggcccggctc cgcgccgcca ccaccgccac cgctcagcgc
      181 cgccatggtg ccaacttttt tgtacaaagt tggcattata aaaaagcatt gctcatcaat
      241 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttggg gcccgagctt
      301 aagactggcc gtcgttttac aacgtcgtga ctgggaaaac atccatgcta gcgttaacgc
      361 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa gactgggcct
      421 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
      481 cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
      541 ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
      601 aaactcttcc tggctagcgg tacgcgtatt aattgcgttg cgctcactgc ccgctttcca
      661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1321 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgta
     1561 actcacgtta agggattttg gtcatgggtg gctcgagggt tatttgccga ctaccttggt
     1621 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     1681 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     1741 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     1801 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     1861 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     1921 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     1981 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     2041 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     2101 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     2161 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     2221 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     2281 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     2341 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca taatgtttaa
     2401 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     2461 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     2521 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     2581 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagct tacgaaccga
     2641 acaggcttat gtccactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     2701 aaccttgggt agcagcgaag tcgaggcatt tctgtcctgg ctggtctaga ggaggcatga
     2761 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccagagctg
     2821 ccaggaaaca gctatgacca tgtaatacga ctcactatag gggatatcag ctggatggca
     2881 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg ataagcaatg
     2941 ctttcttata atgccaactt tgtacaagaa agttggcaat cagtggacag gaaacgcacc
     3001 atatccccct gcctggatgg gtgtttttgg agaagcacaa gcatatagac taaaatcctc
     3061 tgtttggaaa ccagcccgat tcaaggaggg ttctgatgca ctgcggtgaa tttttggcaa
     3121 tgagcgggcc agcagctcaa tagaggcgag aatttgggga aagagtggtc tctcatctct
     3181 tttctttttg aggcactctg ccattaatct cttcatggct tttggacagt tactccgtac
     3241 cttactgaga tctggagaca ggtatcctcg tcccaccata aaaattatct ggtccctgtt
     3301 gttgatgttt gaataaggta actgtccagt catcaattca tacagaacaa ttccaaatgc
     3361 atatacatct gactgaaagc tgtatggatt tttatcttgc attctgatga cttctggtgc
     3421 catccacaaa atggatccag acaactgttc aaactgatgg gacccactcc atcgagattt
     3481 cactgtagct agaccaaaat cacctatttt tactgtgagg tcttcatgaa gaaatatatt
     3541 attactcttg aggtctctgt ggatgattga cttggcgtgt aagtaatcca tgccctgtgc
     3601 agtctgtcgt gcaatatcta taagtttgat catctcaaat ttggtctcaa tgatatggag
     3661 atggtgatac aagctggagc cctcacacca ctgggtaaca atagccagtt gtggctttgt
     3721 ggaatagccc atgaagagta ggatattcac atgtcgtgtt ttcctgagta ctcctacttc
     3781 atttttgaag gcttgtaact gctgaggtgt aggtgctgtc acattcaaca ttttcactgc
     3841 cacatcacca tgccactttc ccttgtagac tgttacaaat gatccagatc caattctttg
     3901 tcccactgta atctgcccat caggaatctc ccaatcatca ctcgagtccc gtctaccaag
     3961 tgttttcatt cgattcctgt cttctgagga tgaagatgac ttcctttctc gctgaggtcc
     4021 tggagatttc tgtaaggctt tcacgttagt tagtgagcca ggtaatgagg cagggggggt
     4081 agcagacaaa cctgtggttg atcctccatc accacgaaat ccttggtctc taatcaagtc
     4141 atcaatattg acaggttcta ttgtgtttat atgcacattg ggagctgatg aggatcggtc
     4201 tcgttgccca aattgatttc gatgatcttc atctgctggt cggaagggct gtggaattgg
     4261 aatggatttt gaaggagacg gactggtgag aatttggggc ccaatagagt ccgaggcggg
     4321 tgcggaaggg gatgatccag atgttagggc agtctctgct aaggacgcct cttcctgtgg
     4381 tattgggtgg tgttcaaaga acttggagac aaacagcaaa tcaagttggt cataattaac
     4441 acacatcagt ggaacttctg tactacaacg ctggtgaaat ttataaccac atgtttgaca
     4501 gcggaaaccc tggaaaagca gctttcgaca aaagtcacaa aatgctaagg tgaaaaacgt
     4561 ttttcgtaca aagttgtgtg ttgtaagtgg aacattctcc aacacttcca catgcaattc
     4621 ttctccagta agccaggaaa tatcagtgtc ccaaccaatt ggtttcttct ctccatcctg
     4681 aattctgtaa acagcacagc actctgggat tagacctctc atcatcagtg ctttctttag
     4741 actgtctcgg actgtaactc cacaccttgc aggtaccact gtcctctgtt tgttgggcag
     4801 gaagactcta acgataggtt tttgtggtga cttggggttg ctccgtgcca catctgtggg
     4861 attttgaaaa actgaaagag atgaaggtag cactgaaagg ctagaagagg aagaagatgt
     4921 aacggtatcc attgatgcag agctagaaac agaaaaatca gttccgttcc ccagagattc
     4981 caataactgt tgttctcttt gttggagtgc atctagcttg ctggtgtatt cttcataggc
     5041 ctccagatat attgatggtg gattatgctc cccaccaaat ttgtccaata gggc
//
