LOCUS Exported 5765 bp ds-DNA circular SYN 30-NOV-2021 DEFINITION Template for in vitro transcription of human MLH1dn. ACCESSION . VERSION . KEYWORDS pT7-hMLH1dn for IVT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5765) AUTHORS Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR TITLE Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. JOURNAL Cell. 2021 Oct 9. pii: S0092-8674(21)01065-5. doi: 10.1016/j.cell.2021.09.018. PUBMED 34653350 REFERENCE 2 (bases 1 to 5765) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 30, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5765 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(498..1358) /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 1121..1140 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1359..1463) /gene="bla" /label=AmpR promoter primer_bind complement(1542..1561) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 1733..2112 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2113..2316 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2267..2287 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind complement(4801..4818) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4807..5031 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(5102..5118) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5102..5118) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5115..5137) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5126..5142 /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)." promoter complement(5150..5180) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5195..5216 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5333..5350) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5504..327) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5584..5603) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 61 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 361 actcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 1021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 1081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 1141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 1201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 1261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 1321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 1381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 1441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga 1501 tcgggagatc gatctcccga tcccctaggg tctactctca gtacaatctg ctctgatgcc 1561 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 1621 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 1681 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 1741 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 1801 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 1861 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 1921 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 1981 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 2041 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 2101 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 2161 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 2221 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 2281 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatctcg 2341 agctcggtac ctaatacgac acactataag gaaataagag agaaaagaag agtaagaaga 2401 aatataagag ccaccatgag cttcgttgct ggagtcatcc ggagactgga cgagacagtg 2461 gtgaacagaa ttgccgccgg cgaggtgatc cagagacctg ccaatgcaat aaaggagatg 2521 atcgagaact gtctggacgc caagtccaca agcattcagg tgatcgtgaa ggagggcgga 2581 ctgaagctga tccagatcca agacaacggc acaggcatca gaaaggaaga tctggacatc 2641 gtgtgtgaac ggttcaccac atctaagctg cagtcttttg aggatctggc ctctatcagt 2701 acctacggct tcagaggcga ggccctggcc agcatcagcc acgtggccca tgtgaccatc 2761 accaccaaaa ccgccgacgg caaatgcgct tatcgcgcta gctacagcga cggcaagctg 2821 aaagccccgc caaagccttg cgccggcaac cagggtacac agataacagt ggaggatctg 2881 ttctacaaca tcgccacccg gagaaaggcc ctgaaaaatc ccagcgagga gtacggcaag 2941 atcctggaag tcgtcggcag atactccgtg cacaacgccg gaatcagctt tagcgtaaag 3001 aagcagggag aaaccgtggc cgatgtgcgc accctgccaa atgccagcac cgtggataac 3061 atcagaagca ttttcggaaa tgccgtgtcc agagaactga tcgagatcgg ctgcgaagat 3121 aagaccctgg cttttaagat gaacggctac atcagcaacg ccaattactc tgtgaagaag 3181 tgcatctttc ttctgttcat caaccacaga ctggtggaaa gcaccagcct gcggaaagcc 3241 atcgagacag tgtacgccgc ctacctgcct aagaacaccc accccttcct gtacctgagc 3301 ctcgagatca gccctcagaa cgtggacgtc aatgtgcatc ctacaaagca cgaggtgcac 3361 ttcctgcacg aggaaagcat cctggaaaga gtgcagcagc acattgagag caagctgctg 3421 ggctctaaca gcagcagaat gtacttcaca cagaccctgc tgcctggcct ggccggcccc 3481 tcaggcgaaa tggttaagtc cacaacctct ctgacctcat ccagcaccag cggttcttcc 3541 gataaggtgt acgcccacca gatggtgcgg accgactctc gggagcagaa gctggacgcc 3601 tttctgcaac ctctgagcaa acctctgagc tctcagcctc aggccatcgt gaccgaggac 3661 aagacagata tctcctccgg ccgtgccaga cagcaggacg aagaaatgct cgagctgcca 3721 gctcctgccg aggtggccgc caagaaccag agcctggagg gagataccac aaagggcacc 3781 agcgaaatga gcgagaagcg gggccctacc tccagcaacc ccagaaaacg gcaccgggag 3841 gacagcgacg tggaaatggt ggaggacgac agcagaaagg aaatgacagc cgcttgtacc 3901 cctagaagaa gaatcatcaa cctgacctcc gtgctgagcc tgcaggagga gatcaacgag 3961 cagggccacg aggtgctgag agagatgctg cacaatcaca gcttcgtggg ctgcgtgaac 4021 cctcaatggg ccctggctca gcatcaaaca aagctgtacc tgctgaacac caccaagctg 4081 agcgaagagc tgttctacca gatcctcatc tacgacttcg ccaacttcgg cgtgctacgc 4141 ctgagcgagc ccgcccctct gtttgacctg gccatgctgg ctctggatag cccagaaagc 4201 ggctggacag aagaggacgg acctaaagag gggctggctg aatacatcgt ggagttcctg 4261 aagaaaaagg ccgagatgct ggccgactac ttttctctgg aaatcgacga ggaaggcaac 4321 ctgatcggcc tgcctctgct gatcgataac tacgtgcctc ccctggaagg cctgcccatc 4381 ttcatcctga gactggctac agaggtgaac tgggacgagg aaaaggaatg cttcgagtct 4441 ctgagcaagg agtgcgccat gttctatagc atcagaaaac agtacatctc tgaagagagc 4501 actctgtctg gccagcagag tgaagtgccc ggaagcatcc ccaacagctg gaagtggacc 4561 gtggaacaca tcgtgtacaa ggccctgcgg agccacattc tccctcctaa gcacttcacc 4621 gaggacggca acatcctgca gctggccaac ctgcccgacc tttataaggt tttctaatta 4681 attaagctgc cttctgcggg gcttgccttc tggccatgcc cttcttctct cccttgcacc 4741 tgtacctctt ggtctttgaa taaagcctga gtaggaagtg agtttaaacc cgctgatcag 4801 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 4861 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgagaaa attgcatcgc 4921 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 4981 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 5041 cggaaagaac cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat 5101 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 5161 ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg 5221 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 5281 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 5341 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 5401 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 5461 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 5521 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 5581 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 5641 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5701 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5761 acgaa //