LOCUS       sequence-201880-        6210 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 6210)
  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 6210)
  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-K0XCK7
FEATURES             Location/Qualifiers
     source          1..6210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     277..299
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     291..308
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     292..308
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3822..3842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3852..3868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3852..3868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3865..3887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3876..3892
                     /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(3900..3906,3907..3924,3925..3930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3945..3966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4304..4892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4384..4403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5063..5854,5855..5923))
                     /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     5686..5705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5924..6028)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6096..6114
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6152..6174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
       61 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
      121 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
      241 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      301 cggccagtgt gctgtctcat cattttggca aagaattcgc ggccgccacc atgaaaaaca
      361 tcttggggct ggacctgggc ttgtcctcta tagggtggtc cgtgatacgc gagaattctg
      421 aagaacagga gctcgtggct atgggttcaa gggtggtatc cctgaccgcg gccgaacttt
      481 cttcctttac tcaaggaaat ggagtcagca tcaattctca gcgaactcaa aaaagaactc
      541 aacggaaggg ttacgaccgg tatcagctta ggaggactct gctgagaaac aagcttgata
      601 ccctggggat gttgcccgac gatagtctgt catacctgcc aaagctccag ctgtgggggc
      661 tgagggcgaa ggctgtaaca cagcgcattg aactcaacga gctcggaagg gtccttttgc
      721 atctcaatca gaagcgcggc tacaagagta taaaatccga cttcagcggt gataaaaaga
      781 tcacagacta cgtgaagacc gtcaaaacac gctatgatga actcaaggaa atgagactga
      841 ccatcggaga actctttttc agacgactta ccgagaacgc atttttccgg tgtaaggaac
      901 aggtctatcc ccgccaggct tacgtggaag agttcgactg tatcatgaat tgtcagagga
      961 agttttatcc cgatatcttg actgatgaga ctatccggtg cattagggat gagatcatct
     1021 actatcagcg cccattgaag tcttgtaagt acctggtgag tcgctgcgaa tttgaaaaaa
     1081 gattttacct gaacgccgcc ggtaagaaaa ctgaagccgg ccccaaggtg tctccccgga
     1141 caagtccatt gtttcaggtg tgtcggctgt gggaatccat caataatatc gtggtgaagg
     1201 acaggcggaa cgagatagtg tttatctccg ctgaacagag ggccgccctg tttgattttc
     1261 tcaacaccca tgaaaagctt aagggttcag acctgcttaa attgctgggt ctttcaaaga
     1321 cctatggtta cagactcgga gagcagttta agactgggat tcagggtaat aagacaaggg
     1381 tggagatcga gagagccctt ggcaactatc ccgacaagaa gcggctgttg cagttcaacc
     1441 tccaggaaga aagctcaagt atggtgaata cggaaaccgg tgaaatcatc cccatgattt
     1501 ccctcagctt tgaacaggaa cctctgtatc ggctttggca cgtcctttat tcaattgatg
     1561 acagagagca gttgcagtca gttttgagac agaagttcgg gatcgatgat gacgaagttc
     1621 tggaacgcct gagcgccatt gacctggtta aagccggctt cgggaataag agtagcaagg
     1681 ccattcggcg catcctcccc tttctgcaat tgggaatgaa ctatgcagag gcttgcgaag
     1741 ccgcagggta taatcacagc aacaactata ccaaagccga aaacgaggca cgggccctgc
     1801 tcgatagact tcctgctatc aaaaaaaacg agcttcgaca gcccgtggtg gagaaaatac
     1861 ttaaccaaat ggtgaacgtg gtgaacgcac ttatggagaa gtacgggagg tttgatgaga
     1921 ttagagtcga gttggctagg gaacttaaac agagtaaaga agaacgctca aacacgtaca
     1981 agagtattaa caagaaccag cgggaaaatg agcagatcgc taaaaggatc gtggaatatg
     2041 gggtgccaac taggtcacgg atccaaaagt ataaaatgtg ggaggagtct aagcattgct
     2101 gcatttattg cggtcagcct gttgatgtgg gggacttcct gcgggggttc gatgttgagg
     2161 tggagcacat cataccaaaa agcctctatt ttgacgacag ttttgcaaat aaggtctgtt
     2221 cctgcagaag ctgcaataaa gaaaagaaca atcggacggc ctacgactac atgaagtcca
     2281 agggggagaa ggcccttagc gactatgtcg agcgagttaa tacaatgtac actaataacc
     2341 agataagcaa gaccaagtgg caaaacttgc tgactccagt ggataaaatt tcaatcgatt
     2401 tcatcgaccg gcagcttcgg gaatctcagt acatcgctcg gaaagctaaa gaaatcttga
     2461 ccagtatttg ctacaacgtt accgccacca gcgggtccgt cacgagcttt ctgagacacg
     2521 tgtgggggtg ggataccgtg ctgcacgatc ttaattttga tcggtacaag aaggtgggcc
     2581 tcacagaagt cattgaagtg aaccatagag ggagtgtgat tagacgggaa cagataaaag
     2641 actggtctaa acgctttgac caccgacacc acgctattga tgctttgact atagcttgta
     2701 ctaaacaagc atacatccag cggctgaata accttcgcgc tgaggaagga ccagacttta
     2761 acaagatgtc cttggaacga tacatccaga gccagcccca cttttctgtg gctcaggtgc
     2821 gcgaggccgt ggacaggatt ctcgtgagct ttcgggcagg aaagcgcgct gtcaccccag
     2881 gaaagagata cataaggaaa aacaggaagc ggataagcgt gcagtccgtg ctgatcccca
     2941 gaggcgccct ctcagaagaa tccgtctatg gagtgatcca tgtctgggaa aaggatgagc
     3001 agggccatgt gatccaaaag caacgcgcag tgatgaaata ccccataaca tcaattaaca
     3061 gagaaatgct ggacaaggag aaggtcgtgg ataagagaat ccaccgcatc cttagcggtc
     3121 gactggccca gtacaatgat aatccaaagg aagcatttgc caaacccgtt tacatcgaca
     3181 aagaatgtcg cattcccatc cgcacggttc gatgcttcgc aaaaccagca attaacaccc
     3241 tggttcctct gaaaaaggac gacaagggta atcctgtggc gtgggtcaac cctggtaata
     3301 accaccatgt ggctatctac cgggacgagg atggcaagta taaagagaga accgttacct
     3361 tctgggaggc agtagataga tgtagagtag gaatccccgc tatcgtgacc cagccagata
     3421 caatttggga caatattctg cagcgcaacg acattagcga aaacgttctg gaatccctcc
     3481 ccgatgttaa gtggcagttt gtgttgagtc tgcagcagaa tgagatgttc attctgggta
     3541 tgaacgagga ggactataga tacgcaatgg atcagcagga ttacgctctg ctgaacaaat
     3601 atttgtacag agtacagaag cttagcaagt ccgattactc tttcaggtac catacagaaa
     3661 cgagcgtgga ggataagtat gacggaaaac ctaacctcaa acttagtatg cagatgggga
     3721 agcttaaaag ggtgagcata aaaagccttc tgggtcttaa ccctcacaaa gttcacatct
     3781 ccgtgctggg cgaaatcaaa gaaatctcta gcagggctga ccccaagaag aagaggaagg
     3841 tgaggtccat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3901 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3961 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4021 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4081 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc gtcttccgct tcctcgctca
     4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6061 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6121 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6181 tcacagcttg tctgtaagcg gatgccggga
//
