LOCUS Exported 2635 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Plasmid expressing a sgRNA to introduce BRCA2 V572I using base editing. ACCESSION . VERSION . KEYWORDS sgRNA BRCA2 V572I SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2635) AUTHORS Billon P, Nambiar TS, Hayward SB, Zafra MP, Schatoff EM, Oshima K, Dunbar A, Breinig M, Park YC, Ryu HS, Tschaharganeh DF, Levine RL, Baer R, Ferrando A, Dow LE, Ciccia A TITLE Detection of Marker-Free Precision Genome Editing and Genetic Variation through the Capture of Genomic Signatures. JOURNAL Cell Rep. 2020 Mar 10;30(10):3280-3295.e6. doi: 10.1016/j.celrep.2020.02.068. PUBMED 32160537 REFERENCE 2 (bases 1 to 2635) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2635 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 82..322 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 350..425 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 439..679 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 708..783 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(886..1474) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(966..985) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1645..2505) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 2268..2287 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2506..2610) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc 61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata 121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta 181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta 241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct 301 ttatatatct tgtggaaagg acgaaacacc gagacgatta atgcgtctcg ttttagagct 361 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc 421 ggtgcttttt ttaagcttga gggcctattt cccatgattc cttcatattt gcatatacga 481 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta 541 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt 601 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta 661 tatatcttgt ggaaaggacg aaacaccggc tacagaattc tgtgtgggtt ttagagctag 721 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg 781 tgcttttttt ctagagaatt cctcgaggta cctctctaca tatgacatgt gagcaaaagg 841 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 901 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 961 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 1021 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 1081 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 1141 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 1201 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 1261 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 1321 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 1381 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 1441 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 1501 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 1561 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 1621 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 1681 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 1741 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 1801 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 1861 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 1921 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 1981 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 2041 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 2101 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 2161 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 2221 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 2281 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 2341 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 2401 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 2461 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 2521 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 2581 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacct //