LOCUS Exported 2636 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION B52 plasmid expressing PARP4 sgSTOP (cloned in BbsI site) and containing an empty sgRNA-expression cassette (use BsmBI for cloning). ACCESSION . VERSION . KEYWORDS B52 + PARP4 sgSTOP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2636) AUTHORS Billon P, Bryant EE, Joseph SA, Nambiar TS, Hayward SB, Rothstein R, Ciccia A TITLE CRISPR-Mediated Base Editing Enables Efficient Disruption of Eukaryotic Genes through Induction of STOP Codons. JOURNAL Mol Cell. 2017 Sep 4. pii: S1097-2765(17)30605-6. doi: 10.1016/j.molcel.2017.08.008. PUBMED 28890334 REFERENCE 2 (bases 1 to 2636) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2636 /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 709..784 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(887..1475) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(967..986) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1646..2506) /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 2269..2288 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2507..2611) /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 aaacaccgat ccagtcagca caagtgccgt tttagagcta 721 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg 781 gtgctttttt tctagagaat tcctcgaggt acctctctac atatgacatg tgagcaaaag 841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 1021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 1081 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 1141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 1201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 1261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 1321 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 1381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 1441 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 1501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 1561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 1621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 1681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 1741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 1801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 1861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 1921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 1981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 2041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 2101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 2161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 2221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 2281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 2341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 2401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 2461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 2521 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 2581 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct //