LOCUS Exported 3320 bp ds-DNA circular SYN 03-JUN-2021 DEFINITION pegRNA used for installation (via C•G-to-T•A) of the oncogenic S45F in Ctnnb1 in mouse liver.. ACCESSION . VERSION . KEYWORDS pegCTNNB1 S45F SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3320) AUTHORS Liu P, Liang S, Zheng C, Mintzer E, Zhao YG, Ponnienselvan K, Mir A, Sontheimer EJ, Gao G, Flotte TR, Wolfe SA, Xue W TITLE Improved prime editors enable pathogenic allele correction and cancer modelling in adult mice JOURNAL BioRxiv REFERENCE 2 (bases 1 to 3320) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 3, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3320 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 21..125 /gene="bla" /label=AmpR promoter CDS 126..986 /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 complement(344..363) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1157..1745 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1646..1665 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1899..1916 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2033..2054 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2069..2099 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2107..2123 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2112..2134 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2131..2147 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2131..2147 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2166..2186 /label=T3 /note="T3 promoter, forward primer" promoter 2168..2186 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2233..2473 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2233..2253 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2404..2423 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 2503..2578 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(2632..2648) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2633..2649) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(2673..2692) /label=T7 /note="T7 promoter, forward primer" promoter complement(2674..2692) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2702..2719) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2702..2718) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2711..2733) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2860..3315 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2947..2966) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3157..3178 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga 2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt 2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca 2161 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc 2221 gcccccttca ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag 2281 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat 2341 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa 2401 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc 2461 ttgtggaaag gacgaaacac cgagggttgc ccttgccact cagttttaga gctagaaata 2521 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtgggaccga gtcggtccgc 2581 tcctttcctg agtggcaagg gcaatttttt tgaattcgat atcaagctta tcgataccgt 2641 cgacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat tacgcgcgct 2701 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 2761 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 2821 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat 2881 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 2941 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 3001 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 3061 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 3121 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 3181 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 3241 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 3301 taacgcttac aatttaggtg //