LOCUS Exported 3495 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing TNF , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_TNF_WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3495) 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 3495) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3495 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1374..1473) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1490..1509) /label=T7 /note="T7 promoter, forward primer" promoter complement(1491..1509) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1514..1530) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1514..1530) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1961..2752 /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 2845..3433 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3334..3353 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agttggcacc atgagcactg aaagcatgat ccgggacgtg gagctggccg aggaggcgct 721 ccccaagaag acaggggggc cccagggctc caggcggtgc ttgttcctca gcctcttctc 781 cttcctgatc gtggcaggcg ccaccacgct cttctgcctg ctgcactttg gagtgatcgg 841 cccccagagg gaagagttcc ccagggacct ctctctaatc agccctctgg cccaggcagt 901 cagatcatct tctcgaaccc cgagtgacaa gcctgtagcc catgttgtag caaaccctca 961 agctgagggg cagctccagt ggctgaaccg ccgggccaat gccctcctgg ccaatggcgt 1021 ggagctgaga gataaccagc tggtggtgcc atcagagggc ctgtacctca tctactccca 1081 ggtcctcttc aagggccaag gctgcccctc cacccatgtg ctcctcaccc acaccatcag 1141 ccgcatcgcc gtctcctacc agaccaaggt caacctcctc tctgccatca agagcccctg 1201 ccagagggag accccagagg gggctgaggc caagccctgg tatgagccca tctatctggg 1261 aggggtcttc cagctggaga agggtgaccg actcagcgct gagatcaatc ggcccgacta 1321 tctcgacttt gccgagtctg ggcaggtcta ctttgggatc attgccctgt gattgccaac 1381 tttcttgtac aaagttgtca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca 1441 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga 1501 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg 1561 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag 1621 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat 1681 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg 1741 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt 1801 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc 1861 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt 1921 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc 1981 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac 2041 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga 2101 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat 2161 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga 2221 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact 2281 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat 2341 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg 2401 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa 2461 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt 2521 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt 2581 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc 2641 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga 2701 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag 2761 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc 2821 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 2881 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 2941 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 3001 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 3061 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 3121 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 3181 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 3241 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 3301 cggaacagga gagcgcacga gggaacttcc agggggaaac gcctggtatc tttatagtcc 3361 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 3421 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 3481 ttttgctcac atgtt //