LOCUS Exported 2635 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Plasmid expressing a sgRNA to introduce BRCA1 E638K using base editing. ACCESSION . VERSION . KEYWORDS sgRNA BRCA1 E638K 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 51..291 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 319..394 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 408..648 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 677..752 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(855..1443) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(935..954) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1614..2474) /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 2237..2256 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2475..2579) /gene="bla" /label=AmpR promoter ORIGIN 1 taaaggaacc aattcagtcg actggatccg gtaccaaggt cgggcaggaa gagggcctat 61 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa 121 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat 181 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg 241 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg 301 agacgattaa tgcgtctcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg 361 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt taagcttgag ggcctatttc 421 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta 481 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc 541 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa 601 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcaa 661 ttcagtacaa ttaggtgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt 721 atcaacttga aaaagtggca ccgagtcggt gctttttttc tagagaattc ctcgaggtac 781 ctctctacat atgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 841 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 901 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 961 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 1021 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 1081 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 1141 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 1201 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 1261 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 1321 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 1381 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 1441 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 1501 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 1561 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 1621 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 1681 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 1741 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 1801 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 1861 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 1921 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 1981 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 2041 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 2101 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 2161 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 2221 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 2281 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 2341 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 2401 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 2461 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 2521 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 2581 acatttcccc gaaaagtgcc acctgacgtc gctagctgta caaaaaagca ggctt //