LOCUS       sequence-164048-        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"
     terminator      complement(87..173)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     222..244
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     236..253
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     237..253
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    269..368
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2673..2772)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2789..2808)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2790..2808)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2813..2829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2813..2829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3260..4051
                     /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      4144..4732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4633..4652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4886..4903
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5062..5089)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
       61 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      121 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      181 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      241 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      301 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      361 agcaggctta atggcggcgc tgagcggtgg cggtggtggc ggcgctgagc cgggccaggc
      421 tctgttcaac ggggacatgg agcccgaggc cggcgccggc gccggcgccg cggcctcttc
      481 ggctgcggac cctgccattc cggaggaggt gtggaatatc aaacaaatga ttaagttgac
      541 acaggaacat atagaggccc tattggacaa atttggtggg gagcataatc caccatcaat
      601 atatctggag gcctatgaag aatacaccag caagctagat gcactccaac aaagagaaca
      661 acagttattg gaatctctgg ggaacggaac tgatttttct gtttctagct ctgcatcaat
      721 ggataccgtt acatcttctt cctcttctag cctttcagtg ctaccttcat ctctttcagt
      781 ttttcaaaat cccacagatg tggcacggag caaccccaag tcaccacaaa aacctatcgt
      841 tagagtcttc ctgcccaaca aacagaggac agtggtacct gcaaggtgtg gagttacagt
      901 ccgagacagt ctaaagaaag cactgatgat gagaggtcta ataccagagt gctgtgctgt
      961 ttacagaatt caggatggag agaagaaacc aattggttgg gacactgata tttcctggct
     1021 tactggagaa gaattgcatg tggaagtgtt ggagaatgtt ccacttacaa cacacaactt
     1081 tgtacgaaaa acgtttttca ccttagcatt ttgtgacttt tgtcgaaagc tgcttttcca
     1141 gggtttccgc tgtcaaacat gtggttataa atttcaccag cgttgtagta cagaagttcc
     1201 actgatgtgt gttaattatg accaacttga tttgctgttt gtctccaagt tctttgaaca
     1261 ccacccaata ccacaggaag aggcgtcctt agcagagact gccctaacat ctggatcatc
     1321 cccttccgca cccgcctcgg actctattgg gccccaaatt ctcaccagtc cgtctccttc
     1381 aaaatccatt ccaattccac agcccttccg accagcagat gaagatcatc gaaatcaatt
     1441 tgggcaacga gaccgatcct catcagctcc caatgtgcat ataaacacaa tagaacctgt
     1501 caatattgat gacttgatta gagaccaagg atttcgtggt gatggaggat caaccacagg
     1561 tttgtctgct accccccctg cctcattacc tggctcacta actaacgtga aagccttaca
     1621 gaaatctcca ggacctcagc gagaaaggaa gtcatcttca tcctcagaag acaggaatcg
     1681 aatgaaaaca cttggtagac gggactcgag tgatgattgg gagattcctg atgggcagat
     1741 tacagtggga caaagaattg gatctggatc atttggaaca gtctacaagg gaaagtggca
     1801 tggtgatgtg gcagtgaaaa tgttgaatgt gacagcacct acacctcagc agttacaagc
     1861 cttcaaaaat gaagtaggag tactcaggaa aacacgacat gtgaatatcc tactcttcat
     1921 gggctattcc acaaagccac aactggctat tgttacccag tggtgtgagg gctccagctt
     1981 gtatcaccat ctccatatca ttgagaccaa atttgagatg atcaaactta tagatattgc
     2041 acgacagact gcacagggca tggattactt acacgccaag tcaatcatcc acagagacct
     2101 caagagtaat aatatatttc ttcatgaaga cctcacagta aaaataggtg attttggtct
     2161 agctacagag aaatctcgat ggagtgggtc ccatcagttt gaacagttgt ctggatccat
     2221 tttgtggatg gcaccagaag tcatcagaat gcaagataaa aatccataca gctttcagtc
     2281 agatgtatat gcatttggga ttgttctgta tgaattgatg actggacagt taccttattc
     2341 aaacatcaac aacagggacc agataatttt tatggtggga cgaggatacc tgtctccaga
     2401 tctcagtaag gtacggagta actgtccaaa agccatgaag agattaatgg cagagtgcct
     2461 caaaaagaaa agagatgaga gaccactctt tccccaaatt ctcgcctcta ttgagctgct
     2521 ggcccgctca ttgccaaaaa ttcaccgcag tgcatcagaa ccctccttga atcgggctgg
     2581 tttccaaaca gaggatttta gtctatatgc ttgtgcttct ccaaaaacac ccatccaggc
     2641 agggggatat ggtgcgtttc ctgtccactg agacccagct ttcttgtaca aagttggcat
     2701 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2761 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2821 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2881 aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     2941 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     3001 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     3061 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     3121 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     3181 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     3241 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     3301 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     3361 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     3421 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     3481 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     3541 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     3601 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     3661 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     3721 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     3781 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     3841 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     3901 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     3961 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     4021 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     4081 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4141 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4201 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4261 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4321 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4381 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4441 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4501 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4561 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4621 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4681 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4741 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4801 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4861 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4921 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4981 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     5041 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg ctta
//
