LOCUS Exported 2309 bp DNA circular SYN 20-DEC-2021 DEFINITION Prime editing in mammalian cells. ACCESSION . VERSION . KEYWORDS pU6-pegRNA-GG-acceptor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2309) TITLE Liu Lab plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 2309) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 5, 2022 from SnapGene Viewer 6.0.0 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" FEATURES Location/Qualifiers source 1..2309 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_feature 250..269 /label=guide misc_RNA 270..345 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 346..375 /label=extension rep_origin complement(479..1067) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(559..578) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1238..2098) /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 1861..1880 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2099..2203) /gene="bla" /label=AmpR promoter ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg gagggagtgg tgcatggtgg ttttagagct agaaatagca agttaaaata 301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttacc tgttgtcaca 361 ccatgcacca ctcccttttt ttaagcttgg gccgctcgag gtacctctct acatatgaca 421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 901 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 1021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 1081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 1141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 1201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 1261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 1321 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 1381 atccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 1441 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 1501 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 1561 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 1621 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 1681 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 1741 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 1801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 1861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 1921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 1981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 2041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 2101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 2161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 2221 tgccacctga cgtcgctagc tgtacaaaaa agcaggcttt aaaggaacca attcagtcga 2281 ctggatccgg taccaaggtc gggcaggaa //