LOCUS       sequence-488115-        5777 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Template for in vitro transcription of mouse MLH1dn.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5777)
  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 5777)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 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 (mouse) for IVT
FEATURES             Location/Qualifiers
     source          1..5777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1215..1234)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1406..1785
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1786..1989
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1940..1960
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2083..2092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4486..4503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4492..4716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4787..4803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4787..4803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4800..4822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4811..4827
                     /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(4835..4841,4842..4859,4860..4865))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4880..4901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5018..5035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5189..5777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5269..5288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
     1201 gggtctactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     1261 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     1321 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     1381 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     1441 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1501 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1561 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1621 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1681 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     1741 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     1801 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     1861 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     1921 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1981 agcagagctg gtttagtgaa ccgtcagatc tcgagctcgg tacctaatac gacacactat
     2041 aaggaaataa gagagaaaag aagagtaaga agaaatataa gagccaccat ggcattcgtg
     2101 gctggtgtca tccgccgtct tgatgagact gtcgtcaaca gaattgccgc aggagaagtc
     2161 atccagcgtc cagccaacgc catcaaggaa atgatcgaaa actgtctgga tgccaagagc
     2221 acaaacatac aagttgtggt gaaggagggt ggcctcaaac tcattcagat ccaagacaac
     2281 ggcacgggaa ttagaaaaga agacctggac attgtgtgcg agcgcttcac aacctctaaa
     2341 ttgcagacat ttgaggacct ggcaagcatc agtacttatg gcttccgagg ggaggctctg
     2401 gcctccataa gccatgtggc tcacgtgacc atcacgacaa aaacggctga tggaaagtgc
     2461 gcataccgtg catcctactc ggatgggaaa ctacaggccc ctccaaaacc ttgtgccggc
     2521 aaccagggca cactcatcac tgttgaagat ttgttctaca acattatcac acggcggaag
     2581 gctttaaaga atccaagtga ggaatatggg aagatccttg aagtggtcgg acgatactct
     2641 atccacaact ctgggatttc cttctctgtg aagaagcaag gtgaaactgt ttctgatgtg
     2701 cgcaccttgc ccaatgctac cactgtggac aatattcgca gcatctttgg gaatgcggtc
     2761 tctcgagagc tcattgaggt tggatgtgag gacaagacac tggctttcaa gatgaatggc
     2821 tacatttcca atgccaacta cagcgttaaa aagtgtatct tcctgctgtt tatcaaccat
     2881 aggctggtgg agtcagctgc actccgaaaa gccatagaga cagtatatgc tgcatacttg
     2941 cccaaaaaca cgcatccatt cctctacctg tcgctagaaa tctcccctca gaacgtcgat
     3001 gtcaatgtcc accccaccaa gcatgaagtt cacttccttc atgaggaatc aatactgcaa
     3061 agagtgcagc aacatataga aagcaaatta ctgggttcca acagttcaag aatgtatttc
     3121 acccagactc ttctgcctgg tctggcaggc cctagtggag aagcagcacg gccgaccact
     3181 ggagtggcct caagcagtac cagtggttct ggagacaaag tgtatgccta ccagatggtc
     3241 aggacagact ctcgggagca gaagctcgac gcttttttac agcctgtgtc aagcctggga
     3301 ccctcccagc cacaagatcc tgccccggta aggggagcca ggacagaagg ctccccagag
     3361 cgggccacga gagaggatga agagatgctg gcgctcccag cgccagctga ggccgcagct
     3421 gaatctgaga atctggagag agagtctctg atggagacct ccgatgcagc ccagaaagct
     3481 gcacccacat catctccagg ctcttcaagg aaaaggcaca gagaagactc tgatgtagag
     3541 atggtcgaaa atgcatcagg gaaagaaatg acagctgcct gctacccgcg taggagaatt
     3601 ataaacctga caagtgtgtt atcgttgcag gaggagatta gtgaaaggtg ccacgaaacc
     3661 ttgagagaga tgctgaggaa tcatagcttt gtgggctgtg tgaaccccca atgggccctg
     3721 gctcagcacc agaccaaact atatttgctt aatactacca agctgtcaga agaactgttc
     3781 tatcagatac tcatttatga ctttgccaac tttggagttt tgcggctgag cgagcccgcg
     3841 ccactctttg acctagccat gcttgctctg gacagccctg aaagtggctg gactgaagat
     3901 gacggcccga aggaagggtt ggctgagtat attgtagaat ttctgaagaa gaaagcagag
     3961 atgctcgccg actacttctc cgtagagatc gacgaggagg gcaatcttat cggattacct
     4021 ctcctcatag actcttatgt gccccctttg gaggggctgc ccattttcat cctccgcctc
     4081 gctactgaag taaactggga tgaggagaag gagtgcttcg agagtcttag taaagagtgt
     4141 gcaatgtttt atagcattag gaagcagtac atcctggaag agtccacact gtctgggcag
     4201 cagtcagaca tgcctggtag caccagcaag ccatggaagt ggactgtgga gcacatcatc
     4261 tacaaggcct tccggtccca cctgcttcct cccaagcact ttaccgagga tgggaatgtg
     4321 ctccagctgg ccaatctccc tgatctgtac aaagtttttt aattaattaa gctgccttct
     4381 gcggggcttg ccttctggcc atgcccttct tctctccctt gcacctgtac ctcttggtct
     4441 ttgaataaag cctgagtagg aagtgagttt aaacccgctg atcagcctcg actgtgcctt
     4501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4561 ccactcccac tgtcctttcc taataaaatg agaaaattgc atcgcattgt ctgagtaggt
     4621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4681 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4741 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4801 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4861 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5761 gaaaaaaagg atctcaa
//
