LOCUS Exported 3492 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway Donor vector containing TNF , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_TNF_p.S157R SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3492) 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 3492) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3492 /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(892..991) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1008..1027) /label=T7 /note="T7 promoter, forward primer" promoter complement(1009..1027) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1032..1048) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1032..1048) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1479..2270 /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 2363..2951 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2852..2871 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3105..3122 /label=L4440 /note="L4440 vector, forward primer" terminator 3281..3308 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3400..3486 /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 aagttggcat gagcactgaa agcatgatcc gggacgtgga gctggccgag gaggcgctcc 241 ccaagaagac aggggggccc cagggctcca ggcggtgctt gttcctcagc ctcttctcct 301 tcctgatcgt ggcaggcgcc accacgctct tctgcctgct gcactttgga gtgatcggcc 361 cccagaggga agagttcccc agggacctct ctctaatcag ccctctggcc caggcagtca 421 gatcatcttc tcgaaccccg agtgacaagc ctgtagccca tgttgtagca aaccctcaag 481 ctgaggggca gctccagtgg ctgaaccgcc gggccaatgc cctcctggcc aatggcgtgg 541 agctgagaga taaccagctg gtggtgccat cagagggcct gtacctcatc tactcccagg 601 tcctcttcaa gggccaaggc tgcccctcca cccatgtgct cctcacccac accatcaggc 661 gcatcgccgt ctcctaccag accaaggtca acctcctctc tgccatcaag agcccctgcc 721 agagggagac cccagagggg gctgaggcca agccctggta tgagcccatc tatctgggag 781 gggtcttcca gctggagaag ggtgaccgac tcagcgctga gatcaatcgg cccgactatc 841 tcgactttgc cgagtctggg caggtctact ttgggatcat tgccctgtga ttgccaactt 901 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg 961 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt 1021 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat 1081 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa 1141 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga 1201 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta 1261 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg 1321 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca 1381 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta 1441 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg 1501 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt 1561 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata 1621 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca 1681 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag 1741 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc 1801 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg 1861 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc 1921 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg 1981 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc 2041 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg 2101 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta 2161 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag 2221 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc 2281 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg 2341 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2401 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2461 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 2521 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2581 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 2641 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 2701 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 2761 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 2821 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 2881 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 2941 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 3001 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 3061 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 3121 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 3181 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 3241 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat 3301 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc 3361 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc 3421 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt 3481 tatttgatgc ct //