LOCUS Exported 3972 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing MAP2K1 , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_MAP2K1_p.S218D_S222D SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3972) 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 3972) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3972 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 43..65 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 57..74 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 58..74 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 90..189 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1372..1471) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1488..1507) /label=T7 /note="T7 promoter, forward primer" promoter complement(1489..1507) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1512..1528) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1512..1528) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1959..2750 /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 2843..3431 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3332..3351 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3585..3602 /label=L4440 /note="L4440 vector, forward primer" terminator 3761..3788 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3880..3966 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga 121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa 181 aagttggcac catgcccaag aagaagccga cgcccatcca gctgaacccg gcccccgacg 241 gctctgcagt taacgggacc agctctgcgg agaccaactt ggaggccttg cagaagaagc 301 tggaggagct agagcttgat gagcagcagc gaaagcgcct tgaggccttt cttacccaga 361 agcagaaggt gggagaactg aaggatgacg actttgagaa gatcagtgag ctgggggctg 421 gcaatggcgg tgtggtgttc aaggtctccc acaagccttc tggcctggtc atggccagaa 481 agctaattca tctggagatc aaacccgcaa tccggaacca gatcataagg gagctgcagg 541 ttctgcatga gtgcaactct ccgtacatcg tgggcttcta tggtgcgttc tacagcgatg 601 gcgagatcag tatctgcatg gagcacatgg atggaggttc tctggatcaa gtcctgaaga 661 aagctggaag aattcctgaa caaattttag gaaaagttag cattgctgta ataaaaggcc 721 tgacatatct gagggagaag cacaagatca tgcacagaga tgtcaagccc tccaacatcc 781 tagtcaactc ccgtggggag atcaagctct gtgactttgg ggtcagcggg cagctcatcg 841 acgacatggc caacgacttc gtgggcacaa ggtcctacat gtcgccagaa agactccagg 901 ggactcatta ctctgtgcag tcagacatct ggagcatggg actgtctctg gtagagatgg 961 cggttgggag gtatcccatc cctcctccag atgccaagga gctggagctg atgtttgggt 1021 gccaggtgga aggagatgcg gctgagaccc cacccaggcc aaggaccccc gggaggcccc 1081 ttagctcata cggaatggac agccgacctc ccatggcaat ttttgagttg ttggattaca 1141 tagtcaacga gcctcctcca aaactgccca gtggagtgtt cagtctggaa tttcaagatt 1201 ttgtgaataa atgcttaata aaaaaccccg cagagagagc agatttgaag caactcatgg 1261 ttcatgcttt tatcaagaga tctgatgctg aggaagtgga ttttgcaggt tggctctgct 1321 ccaccatcgg ccttaaccag cccagcacac caacccatgc tgctggcgtc ttgccaactt 1381 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg 1441 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt 1501 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat 1561 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa 1621 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga 1681 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta 1741 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg 1801 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca 1861 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta 1921 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg 1981 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt 2041 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata 2101 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca 2161 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag 2221 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc 2281 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg 2341 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc 2401 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg 2461 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc 2521 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg 2581 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta 2641 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag 2701 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc 2761 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg 2821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 3001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 3061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 3121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 3181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 3241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 3301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 3361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 3421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 3481 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 3541 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 3601 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 3661 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 3721 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat 3781 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc 3841 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc 3901 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt 3961 tatttgatgc ct //