LOCUS Exported 6558 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing ERBB2,part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_ERBB2_p.T182I SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6558) 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 6558) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6558 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 71..659 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 560..579 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 813..830 /label=L4440 /note="L4440 vector, forward primer" terminator 989..1016 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 1108..1194 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 1243..1265 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1257..1274 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1258..1274 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1290..1389 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(5158..5257) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(5274..5293) /label=T7 /note="T7 promoter, forward primer" promoter complement(5275..5293) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5298..5314) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5298..5314) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5745..6536 /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 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca 61 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 721 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 781 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 841 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 901 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg 961 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt 1021 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt 1081 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa 1141 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct 1201 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 1261 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga 1321 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa 1381 aagttggcat ggagctggcg gccttgtgcc gctgggggct cctcctcgcc ctcttgcccc 1441 ccggagccgc gagcacccaa gtgtgcaccg gcacagacat gaagctgcgg ctccctgcca 1501 gtcccgagac ccacctggac atgctccgcc acctctacca gggctgccag gtggtgcagg 1561 gaaacctgga actcacctac ctgcccacca atgccagcct gtccttcctg caggatatcc 1621 aggaggtgca gggctacgtg ctcatcgctc acaaccaagt gaggcaggtc ccactgcaga 1681 ggctgcggat tgtgcgaggc acccagctct ttgaggacaa ctatgccctg gccgtgctag 1741 acaatggaga cccgctgaac aataccaccc ctgtcacagg ggcctcccca ggaggcctgc 1801 gggagctgca gcttcgaagc ctcacagaga tcttgaaagg aggggtcttg atccagcgga 1861 acccccagct ctgctaccag gacacgattt tgtggaagga catcttccac aagaacaacc 1921 agctggctct catactgata gacaccaacc gctctcgggc ctgccacccc tgttctccga 1981 tgtgtaaggg ctcccgctgc tggggagaga gttctgagga ttgtcagagc ctgacgcgca 2041 ctgtctgtgc cggtggctgt gcccgctgca aggggccact gcccactgac tgctgccatg 2101 agcagtgtgc tgccggctgc acgggcccca agcactctga ctgcctggcc tgcctccact 2161 tcaaccacag tggcatctgt gagctgcact gcccagccct ggtcacctac aacacagaca 2221 cgtttgagtc catgcccaat cccgagggcc ggtatacatt cggcgccagc tgtgtgactg 2281 cctgtcccta caactacctt tctacggacg tgggatcctg caccctcgtc tgccccctgc 2341 acaaccaaga ggtgacagca gaggatggaa cacagcggtg tgagaagtgc agcaagccct 2401 gtgcccgagt gtgctatggt ctgggcatgg agcacttgcg agaggtgagg gcagttacca 2461 gtgccaatat ccaggagttt gctggctgca agaagatctt tgggagcctg gcatttctgc 2521 cggagagctt tgatggggac ccagcctcca acactgcccc gctccagcca gagcagctcc 2581 aagtgtttga gactctggaa gagatcacag gttacctata catctcagca tggccggaca 2641 gcctgcctga cctcagcgtc ttccagaacc tgcaagtaat ccggggacga attctgcaca 2701 atggcgccta ctcgctgacc ctgcaagggc tgggcatcag ctggctgggg ctgcgctcac 2761 tgagggaact gggcagtgga ctggccctca tccaccataa cacccacctc tgcttcgtgc 2821 acacggtgcc ctgggaccag ctctttcgga acccgcacca agctctgctc cacactgcca 2881 accggccaga ggacgagtgt gtgggcgagg gcctggcctg ccaccagctg tgcgcccgag 2941 ggcactgctg gggtccaggg cccacccagt gtgtcaactg cagccagttc cttcggggcc 3001 aggagtgcgt ggaggaatgc cgagtactgc aggggctccc cagggagtat gtgaatgcca 3061 ggcactgttt gccgtgccac cctgagtgtc agccccagaa tggctcagtg acctgttttg 3121 gaccggaggc tgaccagtgt gtggcctgtg cccactataa ggaccctccc ttctgcgtgg 3181 cccgctgccc cagcggtgtg aaacctgacc tctcctacat gcccatctgg aagtttccag 3241 atgaggaggg cgcatgccag ccttgcccca tcaactgcac ccactcctgt gtggacctgg 3301 atgacaaggg ctgccccgcc gagcagagag ccagccctct gacgtccatc atctctgcgg 3361 tggttggcat tctgctggtc gtggtcttgg gggtggtctt tgggatcctc atcaagcgac 3421 ggcagcagaa gatccggaag tacacgatgc ggagactgct gcaggaaacg gagctggtgg 3481 agccgctgac acctagcgga gcgatgccca accaggcgca gatgcggatc ctgaaagaga 3541 cggagctgag gaaggtgaag gtgcttggat ctggcgcttt tggcacagtc tacaagggca 3601 tctggatccc tgatggggag aatgtgaaaa ttccagtggc catcaaagtg ttgagggaaa 3661 acacatcccc caaagccaac aaagaaatct tagacgaagc atacgtgatg gctggtgtgg 3721 gctccccata tgtctcccgc cttctgggca tctgcctgac atccacggtg cagctggtga 3781 cacagcttat gccctatggc tgcctcttag accatgtccg ggaaaaccgc ggacgcctgg 3841 gctcccagga cctgctgaac tggtgtatgc agattgccaa ggggatgagc tacctggagg 3901 atgtgcggct cgtacacagg gacttggccg ctcggaacgt gctggtcaag agtcccaacc 3961 atgtcaaaat tacagacttc gggctggctc ggctgctgga cattgacgag acagagtacc 4021 atgcagatgg gggcaaggtg cccatcaagt ggatggcgct ggagtccatt ctccgccggc 4081 ggttcaccca ccagagtgat gtgtggagtt atggtgtgac tgtgtgggag ctgatgactt 4141 ttggggccaa accttacgat gggatcccag cccgggagat ccctgacctg ctggaaaagg 4201 gggagcggct gccccagccc cccatctgca ccattgatgt ctacatgatc atggtcaaat 4261 gttggatgat tgactctgaa tgtcggccaa gattccggga gttggtgtct gaattctccc 4321 gcatggccag ggacccccag cgctttgtgg tcatccagaa tgaggacttg ggcccagcca 4381 gtcccttgga cagcaccttc taccgctcac tgctggagga cgatgacatg ggggacctgg 4441 tggatgctga ggagtatctg gtaccccagc agggcttctt ctgtccagac cctgccccgg 4501 gcgctggggg catggtccac cacaggcacc gcagctcatc taccaggagt ggcggtgggg 4561 acctgacact agggctggag ccctctgaag aggaggcccc caggtctcca ctggcaccct 4621 ccgaaggggc tggctccgat gtatttgatg gtgacctggg aatgggggca gccaaggggc 4681 tgcaaagcct ccccacacat gaccccagcc ctctacagcg gtacagtgag gaccccacag 4741 tacccctgcc ctctgagact gatggctacg ttgcccccct gacctgcagc ccccagcctg 4801 aatatgtgaa ccagccagat gttcggcccc agcccccttc gccccgagag ggccctctgc 4861 ctgctgcccg acctgctggt gccactctgg aaaggcccaa gactctctcc ccagggaaga 4921 atggggtcgt caaagacgtt tttgcctttg ggggtgccgt ggagaacccc gagtacttga 4981 caccccaggg aggagctgcc cctcagcccc accctcctcc tgccttcagc ccagccttcg 5041 acaacctcta ttactgggac caggacccac cagagcgggg ggctccaccc agcaccttca 5101 aagggacacc tacggcagag aacccagagt acctgggtct ggacgtgcca gtgtgattgc 5161 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg 5221 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata 5281 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc 5341 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg 5401 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac 5461 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt 5521 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg 5581 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg 5641 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg 5701 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga 5761 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac 5821 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca 5881 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt 5941 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg 6001 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg 6061 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca 6121 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg 6181 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc 6241 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg 6301 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg 6361 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg 6421 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt 6481 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct 6541 cgagccaccc atgaccaa //