LOCUS       sequence-201802-        6171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 coding gene template with optimized sequence for human codon 
            usage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6171)
  AUTHORS   Cho N, Seo HN, Ryu T, Kwon E, Huh S, Noh J, Yeom H, Hwang B, Ha H, 
            Lee JH, Kwon S, Bang D
  TITLE     High-throughput construction of multiple cas9 gene variants via 
            assembly of high-depth tiled and sequence-verified oligonucleotides.
  JOURNAL   Nucleic Acids Res. 2018 May 18;46(9):e55. doi: 10.1093/nar/gky112.
  PUBMED    29529247
REFERENCE   2  (bases 1 to 6171)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2018-05-18"; volume: "46(9):e55. doi"; pages: " 
            10.1093/nar/gky112"
COMMENT     Sequence Label: pUC19-C7G697
FEATURES             Location/Qualifiers
     source          1..6171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..112
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     306..328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     320..337
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     321..337
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3812..3832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3842..3858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3842..3858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3855..3877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3866..3882
                     /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(3890..3896,3897..3914,3915..3920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3935..3956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4294..4882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4374..4393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5053..5844,5845..5913))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTTFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5676..5695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5914..6018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6086..6104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6142..6164)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
       61 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      121 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
      181 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      241 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      301 gttttcccag tcacgacgtt gtaaaacgac ggccagtgtg ctgtctcatc attttggcaa
      361 agaattcgcg gccgccacca tgagagagaa tggatccgac gagcggaggc ggaatatgga
      421 cgagaaaatg gactaccgga taggacttga tattggaata gcatctgtgg ggtgggccgt
      481 gttgcagaat aactccgacg atgaaccggt aagaatcgta gatttggggg tgagaatttt
      541 cgacacagca gaaatcccaa agaccggcga atccctggca ggaccccggc gcgcagcgcg
      601 caccactcga aggagactga gacgccgaaa acatagactt gacaggatca aatggctgtt
      661 cgaaaaccag gggctgatca acatcgacga tttcctgaag agatataaca tggcaggcct
      721 ccctgacgtg tatcagctga ggtacgaagc tctggaccga aagctgaccg atgaggaact
      781 ggcccaagtc cttctgcaca tcgccaagca tcgggggttt aggtccacac gaaaagcaga
      841 aacagctgct aaagaaaatg gcgcagtgct gaaggccaca gatgaaaacc agaaacgcat
      901 gcaagagaag gggtaccgga ctgtgggaga aatgatttat ctcgacgaag cctttcgcac
      961 ggggtgctct tggtccgaga aaggctacat ccttacccca cggaacaaag cggagaatta
     1021 tcagcatacc atgctgaggg caatgctcgt tgaggaggta aaggaaatct tctcctctca
     1081 gaggcgattg ggtaacgaaa aggccacaga agagctcgaa gaaaagtacc tcgaaatcat
     1141 gacctcacag aggtcttttg atctgggccc tggcatgcag cctgacggta agccctcacc
     1201 gtacgccatg gaagggttct cagatagggt cggaaagtgc accttcctgg gggaccaggg
     1261 cgagctgaga ggcgcaaagg gaacatacac tgctgaatac ttcgtggccc tgcagaaaat
     1321 taaccacacc aaactcgtta accaggacgg agagacaaga aattttactg aggaggaacg
     1381 gcgggcgctc actttgcttc tctttactca aaaggaagtg aaatatgccg cggtgcggaa
     1441 gaagctgggt ctgcccgaag acatcctctt ttacaatctc aactataaga aggccgccac
     1501 taaagaagag cagcagaaag agaaccaaaa tacagaaaag gctaagttca tcggaatgcc
     1561 ctattatcat gattataaga aatgcctgga ggagcgggta aagtacctga cagagaatga
     1621 agtgcgcgac ctgtttgatg agatcgggat gatcctgacc tgctacaaga atgatgactc
     1681 ccggactgaa cgacttgcta agctgggttt ggtgcctatc gagatggagg gattgctggc
     1741 ctatacccca acgaaatttc aacacctgtc tatgaaggcg atgagaaaca ttattccatt
     1801 tctcgagaag ggtatgacct acgacaaggc ctgtgaagag gccggatacg atttcaaggc
     1861 agatagcaaa ggaactaaac aaaagctcct gactggggaa aacgtaaatc agaccatcaa
     1921 tgagatcact aaccccgtgg tcaagaggtc cgtgtctcag actgtgaagg tgatcaacgc
     1981 cattattcgg acatatgggt ccccgcaagc cattaacata gagctggcgc gggagatgtc
     2041 taagactttt gaggagagga gaaagatcaa gggtgatatg gagaaaaggc agaagaacaa
     2101 cgaggacgtg aagaaacaga ttcaggaact ggggaagctg tctccaaccg gacaggacat
     2161 tctgaagtac cggctctggc aagaacagca gggcatctgt atgtatagcg gaaagaccat
     2221 ccccttggaa gagctcttta agccaggata tgacattgat cacatcctgc cttactccat
     2281 taccttcgac gactcattcc gcaacaaggt actcgtgaca agccaggaga acaggcaaaa
     2341 agggaacagg acaccatatg aatacatggg gaatgacgaa cagcgctgga atgagtttga
     2401 aaccagggtt aagactacca ttagagacta caagaagcag caaaagctgc tgaagaagca
     2461 ttttagcgag gaggaaaggt ccgaattcaa ggagcggaac cttacagaca caaagtacat
     2521 taccaccgtg atctataata tgatcagaca aaatctggaa atggctcccc tcaaccgccc
     2581 tgaaaagaag aagcaggttc gcgccgttaa tggagccatc acagcttacc tgaggaaaag
     2641 atgggggctg ccacaaaaga atcgggaaac agatacacat cacgctatgg atgcagtggt
     2701 tatcgcctgc tgtacggatg gtatgatcca gaagattagc cgctacacta aggtacgaga
     2761 gaggtgctac agtaaaggga ctgagtttgt ggacgcagaa accggtgaaa ttttccggcc
     2821 agaggattat tctagggcag agtgggatga aatattcgga gtgcatatcc caaagccatg
     2881 ggaaactttt agagccgaac ttgacgtcag aatgggagac gaccccaagg gtttcctgga
     2941 tacacatagt gacgttgccc tggagctcga ctacccagag tatatttatg agaacctgcg
     3001 acctatcttc gtgtccagaa tgccgaatca caaggtgacc ggggcagccc atgcagatac
     3061 catcagaagc ccgcggcact ttaaggatga ggggatagtt ctgaccaaga ctgctctgac
     3121 agacctgaag ttggataaag acggtgaaat tgatgggtat tacaacccgc agtctgacct
     3181 cctgctttat gaggccttga agaagcaact tctcctgtat ggtaatgacg ccaagaaggc
     3241 tttcgcccag gacttccaca agccgaaagc ggacggcaca gaggggcctg tcgtccggaa
     3301 agtaaagatc cagaagaagc agactatggg ggttttcgtt gatagtggca acgggatcgc
     3361 agagaatggc ggaatggtta gaatcgatgt gtttcgagtg aacggcaagt actacttcgt
     3421 gcccgtgtac accgccgacg tggtgaagaa agtactgccc aatcgcgcct ccacggctca
     3481 caagccttat ggagagtgga aagtgatgga ggataaagat ttcctgtttt ctctgtactc
     3541 ccgggacttg atacacatta aatcaaagaa ggacataccc attaagatgg tgaatggcgg
     3601 gatggaaggg atcaaggaga cttacgcgta ctatatagga gcagatatca gcgcagcgaa
     3661 catccaaggc atcgcccacg actcacggta caaattccgg gggttgggga tccagagcct
     3721 cgatgtcctg gaaaagtgcc agatcgacgt gttgggccac gtcagtgtgg tgcggtccga
     3781 gaaacgcatg gggttcagca gcagggctga ccccaagaag aagaggaagg tgaggtccat
     3841 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3901 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3961 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4081 ctgactcgct gcgctcggtc gttcggctgc gtcttccgct tcctcgctca ctgactcgct
     4141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4201 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5461 gtgcaaaaaa gtggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6061 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg t
//
