LOCUS       sequence-168429-        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.N581S
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      40..628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     529..548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     782..799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(958..985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1077..1163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1212..1234
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1226..1243
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1227..1243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1259..1358
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1359..1368
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3663..3762)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3779..3798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3780..3798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3803..3819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3803..3819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4250..5041
                     /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 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
       61 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      781 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      901 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
      961 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     1021 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     1081 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     1141 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     1201 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
     1261 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
     1321 cttttttata atgccaactt tgtacaaaaa agttggcacc atggcggcgc tgagcggtgg
     1381 cggtggtggc ggcgcggagc cgggccaggc tctgttcaac ggggacatgg agcccgaggc
     1441 cggcgccggc gccggcgccg cggcctcttc ggctgcggac cctgccattc cggaggaggt
     1501 gtggaatatc aaacaaatga ttaagttgac acaggaacat atagaggccc tattggacaa
     1561 atttggtggg gagcataatc caccatcaat atatctggag gcctatgaag aatacaccag
     1621 caagctagat gcactccaac aaagagaaca acagttattg gaatctctgg ggaacggaac
     1681 tgatttttct gtttctagct ctgcatcaat ggataccgtt acatcttctt cctcttctag
     1741 cctttcagtg ctaccttcat ctctttcagt ttttcaaaat cccacagatg tggcacggag
     1801 caaccccaag tcaccacaaa aacctatcgt tagagtcttc ctgcccaaca aacagaggac
     1861 agtggtacct gcaaggtgtg gagttacagt ccgagacagt ctaaagaaag cactgatgat
     1921 gagaggtcta atcccagagt gctgtgctgt ttacagaatt caggatggag agaagaaacc
     1981 aattggttgg gacactgata tttcctggct tactggagaa gaattgcatg tggaagtgtt
     2041 ggagaatgtt ccacttacaa cacacaactt tgtacgaaaa acgtttttca ccttagcatt
     2101 ttgtgacttt tgtcgaaagc tgcttttcca gggtttccgc tgtcaaacat gtggttataa
     2161 atttcaccag cgttgtagta cagaagttcc actgatgtgt gttaattatg accaacttga
     2221 tttgctgttt gtctccaagt tctttgaaca ccacccaata ccacaggaag aggcgtcctt
     2281 agcagagact gccctaacat ctggatcatc cccttccgca cccgcctcgg actctattgg
     2341 gccccaaatt ctcaccagtc cgtctccttc aaaatccatt ccaattccac agcccttccg
     2401 accagcagat gaagatcatc gaaatcaatt tgggcaacga gaccgatcct catcagctcc
     2461 caatgtgcat ataaacacaa tagaacctgt caatattgat gacttgatta gagaccaagg
     2521 atttcgtggt gatggaggat caaccacagg tttgtctgct accccccctg cctcattacc
     2581 tggctcacta actaacgtga aagccttaca gaaatctcca ggacctcagc gagaaaggaa
     2641 gtcatcttca tcctcagaag acaggaatcg aatgaaaaca cttggtagac gggactcgag
     2701 tgatgattgg gagattcctg atgggcagat tacagtggga caaagaattg gatctggatc
     2761 atttggaaca gtctacaagg gaaagtggca tggtgatgtg gcagtgaaaa tgttgaatgt
     2821 gacagcacct acacctcagc agttacaagc cttcaaaaat gaagtaggag tactcaggaa
     2881 aacacgacat gtgaatatcc tactcttcat gggctattcc acaaagccac aactggctat
     2941 tgttacccag tggtgtgagg gctccagctt gtatcaccat ctccatatca ttgagaccaa
     3001 atttgagatg atcaaactta tagatattgc acgacagact gcacagggca tggattactt
     3061 acacgccaag tcaatcatcc acagagacct caagagtaat agtatatttc ttcatgaaga
     3121 cctcacagta aaaataggtg attttggtct agctacagtg aaatctcgat ggagtgggtc
     3181 ccatcagttt gaacagttgt ctggatccat tttgtggatg gcaccagaag tcatcagaat
     3241 gcaagataaa aatccataca gctttcagtc agatgtatat gcatttggaa ttgttctgta
     3301 tgaattgatg actggacagt taccttattc aaacatcaac aacagggacc agataatttt
     3361 tatggtggga cgaggatacc tgtctccaga tctcagtaag gtacggagta actgtccaaa
     3421 agccatgaag agattaatgg cagagtgcct caaaaagaaa agagatgaga gaccactctt
     3481 tccccaaatt ctcgcctcta ttgagctgct ggcccgctca ttgccaaaaa ttcaccgcag
     3541 tgcatcagaa ccctccttga atcgggctgg tttccaaaca gaggatttta gtctatatgc
     3601 ttgtgcttct ccaaaaacac ccatccaggc agggggatat ggtgcgtttc ctgtccactg
     3661 attgccaact ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     3721 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     3781 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     3841 aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag
     3901 ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg
     3961 gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg
     4021 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     4081 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     4141 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     4201 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc
     4261 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     4321 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     4381 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     4441 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     4501 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     4561 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     4621 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     4681 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     4741 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     4801 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     4861 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     4921 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     4981 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     5041 accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt tcca
//
