LOCUS       sequence-168690-        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.G466E
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     761..778
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(937..964)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1056..1142)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1191..1213
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1205..1222
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1206..1222
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1238..1337
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1338..1347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3642..3741)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3758..3777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3759..3777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3782..3798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3782..3798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4229..5020
                     /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 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      781 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      841 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
      901 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
      961 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     1021 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     1081 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     1141 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
     1201 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
     1261 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
     1321 gtacaaaaaa gttggcacca tggcggcgct gagcggtggc ggtggtggcg gcgcggagcc
     1381 gggccaggct ctgttcaacg gggacatgga gcccgaggcc ggcgccggcg ccggcgccgc
     1441 ggcctcttcg gctgcggacc ctgccattcc ggaggaggtg tggaatatca aacaaatgat
     1501 taagttgaca caggaacata tagaggccct attggacaaa tttggtgggg agcataatcc
     1561 accatcaata tatctggagg cctatgaaga atacaccagc aagctagatg cactccaaca
     1621 aagagaacaa cagttattgg aatctctggg gaacggaact gatttttctg tttctagctc
     1681 tgcatcaatg gataccgtta catcttcttc ctcttctagc ctttcagtgc taccttcatc
     1741 tctttcagtt tttcaaaatc ccacagatgt ggcacggagc aaccccaagt caccacaaaa
     1801 acctatcgtt agagtcttcc tgcccaacaa acagaggaca gtggtacctg caaggtgtgg
     1861 agttacagtc cgagacagtc taaagaaagc actgatgatg agaggtctaa tcccagagtg
     1921 ctgtgctgtt tacagaattc aggatggaga gaagaaacca attggttggg acactgatat
     1981 ttcctggctt actggagaag aattgcatgt ggaagtgttg gagaatgttc cacttacaac
     2041 acacaacttt gtacgaaaaa cgtttttcac cttagcattt tgtgactttt gtcgaaagct
     2101 gcttttccag ggtttccgct gtcaaacatg tggttataaa tttcaccagc gttgtagtac
     2161 agaagttcca ctgatgtgtg ttaattatga ccaacttgat ttgctgtttg tctccaagtt
     2221 ctttgaacac cacccaatac cacaggaaga ggcgtcctta gcagagactg ccctaacatc
     2281 tggatcatcc ccttccgcac ccgcctcgga ctctattggg ccccaaattc tcaccagtcc
     2341 gtctccttca aaatccattc caattccaca gcccttccga ccagcagatg aagatcatcg
     2401 aaatcaattt gggcaacgag accgatcctc atcagctccc aatgtgcata taaacacaat
     2461 agaacctgtc aatattgatg acttgattag agaccaagga tttcgtggtg atggaggatc
     2521 aaccacaggt ttgtctgcta ccccccctgc ctcattacct ggctcactaa ctaacgtgaa
     2581 agccttacag aaatctccag gacctcagcg agaaaggaag tcatcttcat cctcagaaga
     2641 caggaatcga atgaaaacac ttggtagacg ggactcgagt gatgattggg agattcctga
     2701 tgggcagatt acagtgggac aaagaattgg atctgaatca tttggaacag tctacaaggg
     2761 aaagtggcat ggtgatgtgg cagtgaaaat gttgaatgtg acagcaccta cacctcagca
     2821 gttacaagcc ttcaaaaatg aagtaggagt actcaggaaa acacgacatg tgaatatcct
     2881 actcttcatg ggctattcca caaagccaca actggctatt gttacccagt ggtgtgaggg
     2941 ctccagcttg tatcaccatc tccatatcat tgagaccaaa tttgagatga tcaaacttat
     3001 agatattgca cgacagactg cacagggcat ggattactta cacgccaagt caatcatcca
     3061 cagagacctc aagagtaata atatatttct tcatgaagac ctcacagtaa aaataggtga
     3121 ttttggtcta gctacagtga aatctcgatg gagtgggtcc catcagtttg aacagttgtc
     3181 tggatccatt ttgtggatgg caccagaagt catcagaatg caagataaaa atccatacag
     3241 ctttcagtca gatgtatatg catttggaat tgttctgtat gaattgatga ctggacagtt
     3301 accttattca aacatcaaca acagggacca gataattttt atggtgggac gaggatacct
     3361 gtctccagat ctcagtaagg tacggagtaa ctgtccaaaa gccatgaaga gattaatggc
     3421 agagtgcctc aaaaagaaaa gagatgagag accactcttt ccccaaattc tcgcctctat
     3481 tgagctgctg gcccgctcat tgccaaaaat tcaccgcagt gcatcagaac cctccttgaa
     3541 tcgggctggt ttccaaacag aggattttag tctatatgct tgtgcttctc caaaaacacc
     3601 catccaggca gggggatatg gtgcgtttcc tgtccactga ttgccaactt tcttgtacaa
     3661 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     3721 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     3781 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     3841 acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac
     3901 ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     3961 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     4021 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     4081 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     4141 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     4201 cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac
     4261 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     4321 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     4381 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     4441 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     4501 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     4561 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     4621 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     4681 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     4741 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     4801 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     4861 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     4921 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     4981 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     5041 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg taga
//
