LOCUS       sequence-349064-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for in vitro transcription of human MLH1dn.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 28;184(22):5635-5652.e29. doi: 
            10.1016/j.cell.2021.09.018.
  PUBMED    34653350
REFERENCE   2  (bases 1 to 5765)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2021.09"; journalName: "Cell"; date: "2021-10-28- 
            28"; volume: "184"; issue: "22"; pages: "5635-5652.e29"
COMMENT     Sequence Label: pT7-hMLH1dn for IVT
FEATURES             Location/Qualifiers
     source          1..5765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(498..1289,1290..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(join(5150..5156,5157..5174,5175..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(join(5504..5765,1..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
//
