LOCUS       sequence-310184-        5178 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of dCas9-BirA*.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5178)
  AUTHORS   Wang AS, Chen LC, Wu RA, Hao Y, McSwiggen DT, Heckert AB, Richardson
            CD, Gowen BG, Kazane KR, Vu JT, Wyman SK, Shin JJ, Darzacq X, Walter
            JC, Corn JE
  TITLE     The Histone Chaperone FACT Induces Cas9 Multi-turnover Behavior and 
            Modifies Genome Manipulation in Human Cells.
  JOURNAL   Mol Cell. 2020 Jul 16;79(2):221-233.e5. doi: 
            10.1016/j.molcel.2020.06.014. Epub 2020 Jun 29.
  PUBMED    32603710
REFERENCE   2  (bases 1 to 5178)
  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.molcel.2020.06.014"; journalName: "Mol Cell"; date: 
            "2020-07-16- 16"; volume: "79"; issue: "2"; pages: "221-233.e5"
COMMENT     Sequence Label: T7pr-His6-MBP-TEV-dCas9-BirA*
FEATURES             Location/Qualifiers
     source          1..5178
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..24,25..27,28..2514,2515..2517,2518..4101)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1..4101
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4102..4149
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(4186..4533,4534..4536,4537..5145)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             5149..5175
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
       61 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      121 agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac agccgaggcc
      181 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      241 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      301 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      361 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      421 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      481 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      541 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      601 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
      661 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
      721 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
      781 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
      841 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
      901 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
      961 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     1021 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     1081 tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     1141 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     1201 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     1261 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     1321 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     1381 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     1441 gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     1501 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac
     1561 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     1621 ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     1681 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     1741 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     1801 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     1861 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     1921 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     1981 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     2041 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     2101 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     2161 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     2221 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     2281 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     2341 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     2401 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     2461 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggacgctatc
     2521 gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac tcggagcgac
     2581 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     2641 tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga caatctgacc
     2701 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     2761 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     2821 aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct gaagtccaag
     2881 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     2941 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     3001 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     3061 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     3121 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     3181 ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg ggactttgcc
     3241 accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     3301 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa gctgatcgcc
     3361 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     3421 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     3481 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     3541 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     3601 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     3661 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     3721 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     3781 cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     3841 ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag agacaagcct
     3901 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     3961 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     4021 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     4081 ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa
     4141 aagaaaaagg gcggtggtag cggtggcgga agcggtggag gcagtaagga caacaccgtg
     4201 cccctgaagc tgatcgccct gctggccaac ggcgagttcc actctggcga gcagctggga
     4261 gagaccctgg gaatgagcag agccgccatc aacaagcaca tccagacact gagagactgg
     4321 ggagtggacg tgttcaccgt gcctggcaag ggctacagcc tgcctgagcc tatccagctg
     4381 ctgaacgcca agcagatcct gggacagctg gatggcggaa gcgtggccgt gctgcctgtg
     4441 atcgactcca ccaatcagta cctgctggac agaatcggag agctgaagtc cggcgacgcc
     4501 tgcatcgccg agtaccagca ggctggcaga ggaggcagag gacggaagtg gttcagccca
     4561 ttcggagcca acctgtacct gtccatgttc tggagactgg agcagggacc tgctgctgcc
     4621 atcggactga gtctggtgat cggaatcgtg atggccgagg tgctgagaaa gctgggagcc
     4681 gacaaggtga gagtgaagtg gcctaatgac ctgtacctcc aggaccgcaa gctggctggc
     4741 atcctggtgg agctgacagg caagacaggc gatgccgctc agatcgtgat cggagccgga
     4801 atcaacatgg ccatgagaag agtggaggag agcgtggtga accagggctg gatcaccctg
     4861 caggaggctg gcatcaacct ggaccggaac accctggccg ccatgctgat cagagagctg
     4921 agagccgctc tggagctgtt cgagcaggag ggactggctc cttacctgag cagatgggag
     4981 aagctggaca acttcatcaa cagacctgtg aagctgatca tcggcgacaa ggaaatcttc
     5041 ggcatctcca gaggaatcga caagcaggga gctctgctgc tggagcagga cggaatcatc
     5101 aagccctgga tgggcggaga aatctccctg agaagcgcag agaaggctta cccttacgat
     5161 gtaccggatt acgcatag
//
