LOCUS       sequence-161656-        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.G469S
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2137..2236)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2253..2272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2254..2272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2277..2293)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2277..2293)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2724..3515
                     /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      3608..4196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4097..4116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4350..4367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4526..4553)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4645..4731)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4780..4802
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4794..4811
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4795..4811
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4827..4926
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      4927..4936
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tgacacagga acatatagag gccctattgg acaaatttgg tggggagcat aatccaccat
       61 caatatatct ggaggcctat gaagaataca ccagcaagct agatgcactc caacaaagag
      121 aacaacagtt attggaatct ctggggaacg gaactgattt ttctgtttct agctctgcat
      181 caatggatac cgttacatct tcttcctctt ctagcctttc agtgctacct tcatctcttt
      241 cagtttttca aaatcccaca gatgtggcac ggagcaaccc caagtcacca caaaaaccta
      301 tcgttagagt cttcctgccc aacaaacaga ggacagtggt acctgcaagg tgtggagtta
      361 cagtccgaga cagtctaaag aaagcactga tgatgagagg tctaatccca gagtgctgtg
      421 ctgtttacag aattcaggat ggagagaaga aaccaattgg ttgggacact gatatttcct
      481 ggcttactgg agaagaattg catgtggaag tgttggagaa tgttccactt acaacacaca
      541 actttgtacg aaaaacgttt ttcaccttag cattttgtga cttttgtcga aagctgcttt
      601 tccagggttt ccgctgtcaa acatgtggtt ataaatttca ccagcgttgt agtacagaag
      661 ttccactgat gtgtgttaat tatgaccaac ttgatttgct gtttgtctcc aagttctttg
      721 aacaccaccc aataccacag gaagaggcgt ccttagcaga gactgcccta acatctggat
      781 catccccttc cgcacccgcc tcggactcta ttgggcccca aattctcacc agtccgtctc
      841 cttcaaaatc cattccaatt ccacagccct tccgaccagc agatgaagat catcgaaatc
      901 aatttgggca acgagaccga tcctcatcag ctcccaatgt gcatataaac acaatagaac
      961 ctgtcaatat tgatgacttg attagagacc aaggatttcg tggtgatgga ggatcaacca
     1021 caggtttgtc tgctaccccc cctgcctcat tacctggctc actaactaac gtgaaagcct
     1081 tacagaaatc tccaggacct cagcgagaaa ggaagtcatc ttcatcctca gaagacagga
     1141 atcgaatgaa aacacttggt agacgggact cgagtgatga ttgggagatt cctgatgggc
     1201 agattacagt gggacaaaga attggatctg gatcattttc aacagtctac aagggaaagt
     1261 ggcatggtga tgtggcagtg aaaatgttga atgtgacagc acctacacct cagcagttac
     1321 aagccttcaa aaatgaagta ggagtactca ggaaaacacg acatgtgaat atcctactct
     1381 tcatgggcta ttccacaaag ccacaactgg ctattgttac ccagtggtgt gagggctcca
     1441 gcttgtatca ccatctccat atcattgaga ccaaatttga gatgatcaaa cttatagata
     1501 ttgcacgaca gactgcacag ggcatggatt acttacacgc caagtcaatc atccacagag
     1561 acctcaagag taataatata tttcttcatg aagacctcac agtaaaaata ggtgattttg
     1621 gtctagctac agtgaaatct cgatggagtg ggtcccatca gtttgaacag ttgtctggat
     1681 ccattttgtg gatggcacca gaagtcatca gaatgcaaga taaaaatcca tacagctttc
     1741 agtcagatgt atatgcattt ggaattgttc tgtatgaatt gatgactgga cagttacctt
     1801 attcaaacat caacaacagg gaccagataa tttttatggt gggacgagga tacctgtctc
     1861 cagatctcag taaggtacgg agtaactgtc caaaagccat gaagagatta atggcagagt
     1921 gcctcaaaaa gaaaagagat gagagaccac tctttcccca aattctcgcc tctattgagc
     1981 tgctggcccg ctcattgcca aaaattcacc gcagtgcatc agaaccctcc ttgaatcggg
     2041 ctggtttcca aacagaggat tttagtctat atgcttgtgc ttctccaaaa acacccatcc
     2101 aggcaggggg atatggtgcg tttcctgtcc actgattgcc aactttcttg tacaaagttg
     2161 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     2221 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     2281 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2341 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     2401 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     2461 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     2521 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     2581 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     2641 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     2701 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     2761 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     2821 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     2881 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     2941 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     3001 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     3061 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     3121 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     3181 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     3241 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     3301 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     3361 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     3421 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     3481 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     3541 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3721 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4321 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4381 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4441 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     4501 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     4561 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     4621 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     4681 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     4741 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa
     4801 cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg atagtgacct
     4861 gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg tacaaaaaag
     4921 ttggcaccat ggcggcgctg agcggtggcg gtggtggcgg cgcggagccg ggccaggctc
     4981 tgttcaacgg ggacatggag cccgaggccg gcgccggcgc cggcgccgcg gcctcttcgg
     5041 ctgcggaccc tgccattccg gaggaggtgt ggaatatcaa acaaatgatt aagt
//
