LOCUS       sequence-240616-        5100 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  C to T base editor for actinomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5100)
  AUTHORS   Tong Y, Whitford CM, Robertsen HL, Blin K, Jorgensen TS, Klitgaard 
            AK, Gren T, Jiang X, Weber T, Lee SY
  TITLE     Highly efficient DSB-free base editing for streptomycetes with 
            CRISPR-BEST.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi: 
            10.1073/pnas.1913493116. Epub 2019 Sep 23.
  PUBMED    31548381
REFERENCE   2  (bases 1 to 5100)
  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: 
            "10.1073/pnas.1913493116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-8- 8"; volume: "116"; issue: "41"; pages: 
            "20366-20375"
COMMENT     Sequence Label: pCRISPR-cBEST
FEATURES             Location/Qualifiers
     source          1..5100
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..687
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(736..759,760..762,763..4836)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4849..5100
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
ORIGIN
        1 atgtcctccg agaccggccc cgtggccgtg gacccgaccc tgcgtcgccg catcgagccg
       61 cacgagttcg aggtcttctt cgacccgcgc gagctgcgca aggagacctg cctgctgtac
      121 gagatcaact ggggcggccg gcactccatc tggcgccaca cctcgcagaa caccaacaag
      181 cacgtcgagg tcaacttcat cgagaagttc accaccgagc ggtacttctg cccgaacacc
      241 cgctgctcca tcacctggtt cctgtcctgg tccccctgcg gcgagtgctc ccgggccatc
      301 accgagttcc tgtcccgcta cccgcacgtc accctgttca tctacatcgc ccggctgtac
      361 caccacgccg acccgcgcaa ccggcagggc ctgcgcgacc tgatctcctc cggcgtcacc
      421 atccagatca tgaccgagca ggagtccggc tactgctggc ggaacttcgt caactactcc
      481 ccgtccaacg aggcccactg gccccgctac ccgcacctgt gggtccgcct gtacgtcctg
      541 gagctgtact gcatcatcct gggcctgccg ccctgcctga acatcctgcg tcgcaagcag
      601 ccgcagctga ccttcttcac catcgccctc cagtcctgcc actaccagcg cctgccgccc
      661 cacatcctgt gggccaccgg cctgaagtcc ggctccgaga ccccgggcac ctcggagtcc
      721 gccaccccgg agtccgacaa gaagtactcc atcggcctcg cgatcggcac caactccgtg
      781 ggctgggcgg tcatcaccga cgagtacaag gtcccctcca agaagttcaa ggtcctgggc
      841 aacaccgacc ggcactcgat caagaagaac ctgatcggcg ccctgctctt cgacagcggc
      901 gagaccgccg aggcgacccg cctgaagcgg accgcgcgtc gccgctacac ccggcgcaag
      961 aaccgcatct gctacctgca ggaaatcttc tccaacgaga tggccaaggt ggacgactcg
     1021 ttcttccacc gcctggagga gagcttcctg gtggaggagg acaagaagca cgagcgccac
     1081 ccgatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     1141 cacctccgca agaagctggt ggactcgacc gacaaggcgg acctgcggct catctacctg
     1201 gccctcgcgc acatgatcaa gttccgcggc cacttcctca tcgagggcga cctgaacccg
     1261 gacaactccg acgtggacaa gctcttcatc cagctggtgc agacctacaa ccagctgttc
     1321 gaggagaacc ccatcaacgc cagcggcgtg gacgccaagg cgatcctctc cgcgcgcctg
     1381 agcaagtccc ggcgcctgga gaacctcatc gcccagctgc cgggcgagaa gaagaacggc
     1441 ctcttcggca acctgatcgc gctgtcgctc ggcctgaccc ccaacttcaa gagcaacttc
     1501 gacctggccg aggacgcgaa gctccagctg tccaaggaca cctacgacga cgacctggac
     1561 aacctgctcg cccagatcgg cgaccagtac gcggacctct tcctggccgc gaagaacctc
     1621 tcggacgcca tcctgctcag cgacatcctg cgggtcaaca ccgagatcac caaggccccg
     1681 ctgtcggcga gcatgatcaa gcggtacgac gagcaccacc aggacctgac cctgctcaag
     1741 gccctcgtgc gccagcagct gcccgagaag tacaaggaaa tcttcttcga ccagtccaag
     1801 aacggctacg ccggctacat cgacggcggc gcgtcgcagg aggagttcta caagttcatc
     1861 aagccgatcc tggagaagat ggacggcacc gaggagctgc tcgtcaagct gaaccgcgag
     1921 gacctgctcc gcaagcagcg gaccttcgac aacggctcca tcccgcacca gatccacctg
     1981 ggcgagctcc acgccatcct ccggcgccag gaggacttct accccttcct gaaggacaac
     2041 cgcgagaaga tcgagaagat cctgaccttc cgcatcccgt actacgtcgg ccccctggcc
     2101 cgcggcaact cccggttcgc gtggatgacc cggaagtcgg aggagaccat caccccgtgg
     2161 aacttcgagg aggtcgtgga caagggcgcg tccgcgcagt cgttcatcga gcgcatgacc
     2221 aacttcgaca agaacctccc gaacgagaag gtcctgccca agcactccct gctctacgag
     2281 tacttcaccg tgtacaacga gctgaccaag gtcaagtacg tgaccgaggg catgcggaag
     2341 ccggccttcc tgtcgggcga gcagaagaag gcgatcgtgg acctgctctt caagaccaac
     2401 cgcaaggtca ccgtgaagca gctgaaggag gactacttca agaagatcga gtgcttcgac
     2461 tccgtcgaga tcagcggcgt ggaggaccgc ttcaacgcct ccctgggcac ctaccacgac
     2521 ctgctcaaga tcatcaagga caaggacttc ctcgacaacg aggagaacga ggacatcctg
     2581 gaggacatcg tcctcaccct gaccctcttc gaggaccgcg agatgatcga ggagcggctc
     2641 aagacctacg cccacctgtt cgacgacaag gtgatgaagc agctgaagcg tcgccgctac
     2701 accggctggg gccgcctctc ccggaagctg atcaacggca tccgggacaa gcagagcggc
     2761 aagaccatcc tggacttcct caagtccgac ggcttcgcca accgcaactt catgcagctc
     2821 atccacgacg acagcctgac cttcaaggag gacatccaga aggcccaggt ctcgggccag
     2881 ggcgacagcc tccacgagca catcgccaac ctggcgggct ccccggcgat caagaagggc
     2941 atcctccaga ccgtcaaggt cgtggacgag ctggtcaagg tgatgggccg ccacaagccc
     3001 gagaacatcg tgatcgagat ggcccgggag aaccagacca cccagaaggg ccagaagaac
     3061 tcgcgcgagc ggatgaagcg gatcgaggag ggcatcaagg agctcggcag ccagatcctg
     3121 aaggagcacc cggtcgagaa cacccagctg cagaacgaga agctgtacct ctactacctg
     3181 cagaacggcc gcgacatgta cgtggaccag gagctcgaca tcaaccggct gtccgactac
     3241 gacgtggacc acatcgtgcc gcagtccttc ctgaaggacg actcgatcga caacaaggtc
     3301 ctgacccgct cggacaagaa ccggggcaag tccgacaacg tgccctcgga ggaggtcgtg
     3361 aagaagatga agaactactg gcgccagctg ctcaacgcca agctcatcac ccagcgcaag
     3421 ttcgacaacc tgaccaaggc cgagcggggc ggcctgagcg agctcgacaa ggcgggcttc
     3481 atcaagcgcc agctggtcga gacccggcag atcaccaagc acgtggccca gatcctggac
     3541 tcccggatga acaccaagta cgacgagaac gacaagctga tccgcgaggt caaggtgatc
     3601 accctcaaga gcaagctggt ctccgacttc cgcaaggact tccagttcta caaggtccgg
     3661 gagatcaaca actaccacca cgcccacgac gcgtacctga acgccgtcgt gggcaccgcg
     3721 ctgatcaaga agtacccgaa gctggagtcc gagttcgtct acggcgacta caaggtctac
     3781 gacgtgcgca agatgatcgc caagagcgag caggagatcg gcaaggccac cgcgaagtac
     3841 ttcttctact ccaacatcat gaacttcttc aagaccgaga tcaccctggc caacggcgag
     3901 atccgcaagc ggcccctgat cgagaccaac ggcgagaccg gcgagatcgt ctgggacaag
     3961 ggccgcgact tcgccaccgt ccggaaggtg ctgtcgatgc cgcaggtcaa catcgtgaag
     4021 aagaccgagg tgcagaccgg cggcttcagc aaggagtcca tcctccccaa gcgcaacagc
     4081 gacaagctga tcgcccggaa gaaggactgg gacccgaaga agtacggcgg cttcgacagc
     4141 cccaccgtcg cctactccgt gctggtcgtg gcgaaggtcg agaagggcaa gagcaagaag
     4201 ctgaagtccg tgaaggagct gctcggcatc accatcatgg agcgctcctc gttcgagaag
     4261 aacccgatcg acttcctgga ggccaagggc tacaaggagg tcaagaagga cctcatcatc
     4321 aagctgccca agtacagcct gttcgagctg gagaacggcc gcaagcggat gctcgcctcc
     4381 gcgggcgagc tgcagaaggg caacgagctg gccctcccgt cgaagtacgt caacttcctg
     4441 tacctcgcgt cccactacga gaagctgaag ggctcgcccg aggacaacga gcagaagcag
     4501 ctcttcgtgg agcagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     4561 agcaagcgcg tcatcctggc cgacgcgaac ctcgacaagg tgctgtccgc ctacaacaag
     4621 caccgcgaca agccgatccg ggagcaggcg gagaacatca tccacctgtt caccctcacc
     4681 aacctgggcg cccccgccgc gttcaagtac ttcgacacca ccatcgaccg caagcggtac
     4741 acctccacca aggaggtcct cgacgcgacc ctgatccacc agagcatcac cggcctgtac
     4801 gagacccgca tcgacctgtc ccagctcggc ggcgactccg gcggctccac caacctgtcc
     4861 gacatcatcg agaaggagac cggcaagcag ctggtcatcc aggagtccat cctgatgctg
     4921 ccggaggaag tcgaggaagt catcggcaac aagcccgagt ccgacatcct ggtccacacc
     4981 gcctacgacg agtccaccga cgagaacgtc atgctgctga cctcggacgc ccccgagtac
     5041 aagccctggg ccctggtcat ccaggactcc aacggcgaga acaagatcaa gatgctgtga
//
