pYK466

LOCUS       pYK466               9565 bp DNA     circular SYN 11-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9565)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    232..819
                     /label=CMV
     primer_bind     863..882
                     /label=T7
     primer_bind     881..919
                     /label=oYK207_dcas9_F
     CDS             920..5059
                     /label=Translation 748-6546
     CDS             920..5023
                     /label=dCas9 (D839A and N863A nonsense-mutation)
     primer_bind     complement(5036..5085)
                     /label=oYK466_dCas9_R
     primer_bind     complement(5036..5059)
                     /label=oYK464_dCas9_R
     misc_feature    5036..5056
                     /label=NLS (SV40 short)
                     /label=nonstandard type: signal
     terminator      5159..5386
                     /label=bGH PA
     misc_feature    5432..5860
                     /label=f1 origin
     misc_feature    5865..6208
                     /label=SV40 early promoter and origin
     misc_feature    6054..6131
                     /label=SV40 ORI
     CDS             6273..7064
                     /label=Kan/neoR
     misc_feature    7238..7368
                     /label=SV40 early polyadenylation signal
     primer_bind     complement(7404..7424)
                     /label=M13-rev
     misc_binding    complement(7430..7452)
                     /label=LacO
     promoter        complement(7457..7486)
                     /label=lac
     misc_feature    complement(7751..8424)
                     /label=pUC origin
     CDS             complement(8572..9231)
                     /label=AmpR
     promoter        complement(9471..9499)
                     /label=Amp prom
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 ggatcgaacc cttgccacca tggacaagaa gtactccatt gggctcgcta tcggcacaaa
      961 cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt
     1021 tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga
     1081 ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg
     1141 cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga
     1201 tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga
     1261 gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac
     1321 catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat
     1381 ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct
     1441 gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca
     1501 gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc
     1561 taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa
     1621 gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc
     1681 taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga
     1741 tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa
     1801 gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa
     1861 agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt
     1921 gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca
     1981 gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa
     2041 atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa
     2101 cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat
     2161 tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa
     2221 agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc
     2281 cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac
     2341 tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag
     2401 gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct
     2461 gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat
     2521 gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa
     2581 gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg
     2641 tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta
     2701 tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga
     2761 cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga
     2821 acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg
     2881 ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc gagacaagca
     2941 gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc ggaacttcat
     3001 gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag cacaagtttc
     3061 tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc cagctatcaa
     3121 aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca
     3181 taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc agaagggaca
     3241 gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac tggggtccca
     3301 aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc tctacctgta
     3361 ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca atcggctctc
     3421 cgactacgac gtggctgcta tcgtgcccca gtcttttctc aaagatgatt ctattgataa
     3481 taaagtgttg acaagatccg ataaagctag agggaagagt gataacgtcc cctcagaaga
     3541 agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac tgatcacaca
     3601 acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt tggataaagc
     3661 cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg tggcccaaat
     3721 tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc gagaggtgaa
     3781 agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc agttttataa
     3841 ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg cagtggtagg
     3901 cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg gagactataa
     3961 agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca aggccaccgc
     4021 taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta cactggccaa
     4081 tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag aaatcgtgtg
     4141 ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc aggtgaacat
     4201 cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc tcccgaaaag
     4261 gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat acggcggatt
     4321 cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga aagggaagtc
     4381 taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc gatcaagctt
     4441 cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca aaaaagacct
     4501 catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga aacgaatgct
     4561 cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta aatacgttaa
     4621 tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag ataatgagca
     4681 gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg agcaaataag
     4741 cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc tttctgctta
     4801 caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc acttgtttac
     4861 tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca tagacagaaa
     4921 gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt caattacggg
     4981 gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg ctgaccccaa
     5041 gaagaagagg aaggtgtcgt aaggatccac tagtccagtg tggtggaatt ctgcagatat
     5101 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     5161 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5221 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5281 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5341 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     5401 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     5461 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5521 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5581 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5641 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5701 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5761 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5821 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5881 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5941 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6001 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6061 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6121 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6181 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6241 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     6301 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6361 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6421 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6481 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6541 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6601 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6661 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6721 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6781 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6841 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6901 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6961 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     7021 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     7081 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     7141 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     7201 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     7261 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     7321 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7381 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7441 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7501 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7561 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7621 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7681 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7741 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7801 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7861 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7921 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7981 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8041 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8101 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8161 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8221 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8281 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8341 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8401 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8461 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8521 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8581 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8641 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8701 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8761 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8821 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8881 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8941 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9001 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9061 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9121 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9181 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9241 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9301 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9361 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9421 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9481 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9541 tccccgaaaa gtgccacctg acgtc
//