LOCUS       sequence-168611-        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   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_BRAF_p.V600E
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     762..779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(938..965)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1057..1143)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1192..1214
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1206..1223
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1207..1223
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1239..1338
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1339..1348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3643..3742)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3759..3778)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3760..3778)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3783..3799)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3783..3799)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4230..5021
                     /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"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      721 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      781 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      841 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
      901 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
      961 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     1021 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     1081 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     1141 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
     1201 gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac
     1261 tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt
     1321 tgtacaaaaa agttggcacc atggcggcgc tgagcggtgg cggtggtggc ggcgcggagc
     1381 cgggccaggc tctgttcaac ggggacatgg agcccgaggc cggcgccggc gccggcgccg
     1441 cggcctcttc ggctgcggac cctgccattc cggaggaggt gtggaatatc aaacaaatga
     1501 ttaagttgac acaggaacat atagaggccc tattggacaa atttggtggg gagcataatc
     1561 caccatcaat atatctggag gcctatgaag aatacaccag caagctagat gcactccaac
     1621 aaagagaaca acagttattg gaatctctgg ggaacggaac tgatttttct gtttctagct
     1681 ctgcatcaat ggataccgtt acatcttctt cctcttctag cctttcagtg ctaccttcat
     1741 ctctttcagt ttttcaaaat cccacagatg tggcacggag caaccccaag tcaccacaaa
     1801 aacctatcgt tagagtcttc ctgcccaaca aacagaggac agtggtacct gcaaggtgtg
     1861 gagttacagt ccgagacagt ctaaagaaag cactgatgat gagaggtcta atcccagagt
     1921 gctgtgctgt ttacagaatt caggatggag agaagaaacc aattggttgg gacactgata
     1981 tttcctggct tactggagaa gaattgcatg tggaagtgtt ggagaatgtt ccacttacaa
     2041 cacacaactt tgtacgaaaa acgtttttca ccttagcatt ttgtgacttt tgtcgaaagc
     2101 tgcttttcca gggtttccgc tgtcaaacat gtggttataa atttcaccag cgttgtagta
     2161 cagaagttcc actgatgtgt gttaattatg accaacttga tttgctgttt gtctccaagt
     2221 tctttgaaca ccacccaata ccacaggaag aggcgtcctt agcagagact gccctaacat
     2281 ctggatcatc cccttccgca cccgcctcgg actctattgg gccccaaatt ctcaccagtc
     2341 cgtctccttc aaaatccatt ccaattccac agcccttccg accagcagat gaagatcatc
     2401 gaaatcaatt tgggcaacga gaccgatcct catcagctcc caatgtgcat ataaacacaa
     2461 tagaacctgt caatattgat gacttgatta gagaccaagg atttcgtggt gatggaggat
     2521 caaccacagg tttgtctgct accccccctg cctcattacc tggctcacta actaacgtga
     2581 aagccttaca gaaatctcca ggacctcagc gagaaaggaa gtcatcttca tcctcagaag
     2641 acaggaatcg aatgaaaaca cttggtagac gggactcgag tgatgattgg gagattcctg
     2701 atgggcagat tacagtggga caaagaattg gatctggatc atttggaaca gtctacaagg
     2761 gaaagtggca tggtgatgtg gcagtgaaaa tgttgaatgt gacagcacct acacctcagc
     2821 agttacaagc cttcaaaaat gaagtaggag tactcaggaa aacacgacat gtgaatatcc
     2881 tactcttcat gggctattcc acaaagccac aactggctat tgttacccag tggtgtgagg
     2941 gctccagctt gtatcaccat ctccatatca ttgagaccaa atttgagatg atcaaactta
     3001 tagatattgc acgacagact gcacagggca tggattactt acacgccaag tcaatcatcc
     3061 acagagacct caagagtaat aatatatttc ttcatgaaga cctcacagta aaaataggtg
     3121 attttggtct agctacagag aaatctcgat ggagtgggtc ccatcagttt gaacagttgt
     3181 ctggatccat tttgtggatg gcaccagaag tcatcagaat gcaagataaa aatccataca
     3241 gctttcagtc agatgtatat gcatttggaa ttgttctgta tgaattgatg actggacagt
     3301 taccttattc aaacatcaac aacagggacc agataatttt tatggtggga cgaggatacc
     3361 tgtctccaga tctcagtaag gtacggagta actgtccaaa agccatgaag agattaatgg
     3421 cagagtgcct caaaaagaaa agagatgaga gaccactctt tccccaaatt ctcgcctcta
     3481 ttgagctgct ggcccgctca ttgccaaaaa ttcaccgcag tgcatcagaa ccctccttga
     3541 atcgggctgg tttccaaaca gaggatttta gtctatatgc ttgtgcttct ccaaaaacac
     3601 ccatccaggc agggggatat ggtgcgtttc ctgtccactg attgccaact ttcttgtaca
     3661 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     3721 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     3781 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     3841 cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga
     3901 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa
     3961 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
     4021 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     4081 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
     4141 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
     4201 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
     4261 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     4321 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     4381 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     4441 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     4501 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     4561 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     4621 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     4681 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     4741 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     4801 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     4861 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     4921 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     4981 actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac
     5041 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtag
//
