LOCUS Exported 5094 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing BRAF, part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_BRAF_p.V600E 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5094 /organism="synthetic DNA construct" /mol_type="other DNA" 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 938..965 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 //