LOCUS Exported 3846 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing PRKACA , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_PRKACA_WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3846) 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 3846) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3846 /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(1246..1345) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1362..1381) /label=T7 /note="T7 promoter, forward primer" promoter complement(1363..1381) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1386..1402) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1386..1402) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1833..2624 /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 2717..3305 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3206..3225 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3459..3476 /label=L4440 /note="L4440 vector, forward primer" terminator 3635..3662 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3754..3840 /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 catgggcaac gccgccgccg ccaagaaggg cagcgagcag gagagcgtga 241 aagaattctt agccaaagcc aaagaagatt ttcttaaaaa atgggaaagt cccgctcaga 301 acacagccca cttggatcag tttgaacgaa tcaagaccct cggcacgggc tccttcgggc 361 gggtgatgct ggtgaaacac aaggagaccg ggaaccacta tgccatgaag atcctcgaca 421 aacagaaggt ggtgaaactg aaacagatcg aacacaccct gaatgaaaag cgcatcctgc 481 aagctgtcaa ctttccgttc ctcgtcaaac tcgagttctc cttcaaggac aactcaaact 541 tatacatggt catggagtac gtgcccggcg gggagatgtt ctcacaccta cggcggatcg 601 gaaggttcag tgagccccat gcccgtttct acgcggccca gatcgtcctg acctttgagt 661 atctgcactc gctggatctc atctacaggg acctgaagcc ggagaatctg ctcattgacc 721 agcagggcta cattcaggtg acagacttcg gtttcgccaa gcgcgtgaag ggccgcactt 781 ggaccttgtg cggcacccct gagtacctgg cccctgagat tatcctgagc aaaggctaca 841 acaaggccgt ggactggtgg gccctggggg ttcttatcta tgaaatggcc gctggctacc 901 cgcccttctt cgcagaccag cccatccaga tctatgagaa gatcgtctct gggaaggtgc 961 gcttcccttc ccacttcagc tctgacttga aggacctgct gcggaacctc ctgcaggtag 1021 atctcaccaa gcgctttggg aacctcaaga atggggtcaa cgatatcaag aaccacaagt 1081 ggtttgccac aactgactgg attgccatct accagaggaa ggtggaagct cccttcatac 1141 caaagtttaa aggccctggg gatacgagta actttgacga ctatgaggaa gaagaaatcc 1201 gggtctccat caatgagaag tgtggcaagg agttttctga gtttttgcca actttcttgt 1261 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta 1321 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt 1381 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca 1441 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct 1501 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac 1561 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct 1621 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt 1681 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc 1741 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc 1801 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat 1861 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg 1921 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt 1981 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc 2041 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca 2101 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg 2161 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg 2221 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg 2281 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct 2341 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt 2401 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg 2461 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg 2521 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg 2581 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat 2641 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa 2701 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 2761 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 2821 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 2881 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 2941 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 3001 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 3061 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 3121 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 3181 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 3241 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 3301 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 3361 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 3421 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 3481 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 3541 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc 3601 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt 3661 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg 3721 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac 3781 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg 3841 atgcct //