LOCUS       sequence-201895-        6753 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 6753)
  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 6753)
  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-Q927P4
FEATURES             Location/Qualifiers
     source          1..6753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     302..324
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     316..333
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     317..333
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4390..4410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4420..4436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4444..4460
                     /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(4468..4474,4475..4492,4493..4498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4513..4534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4872..5460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4952..4971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5631..6422,6423..6491))
                     /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     6254..6273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6492..6596)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6664..6682
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6720..6742)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
       61 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
      121 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
      181 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
      241 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
      301 tcccagtcac gacgttgtaa aacgacggcc agtgtgctgt ctcatcattt tggcaaagaa
      361 ttcgcggccg ccaccatgaa gaagccatat acaattggac tggatatcgg tactaattct
      421 gtgggatggg cagtgctcac agaccagtac gatctggtta agagaaagat gaaaatcgca
      481 ggcgattctg agaagaagca gatcaaaaaa aatttttggg gagtcaggct tttcgatgag
      541 ggccagactg ctgcagatcg aagaatggcc aggacagcta gacgcaggat tgaacggcga
      601 cggaacagaa tctcttactt gcaagggatc tttgccgagg agatgagcaa gaccgatgcc
      661 aatttctttt gtaggcttag tgattctttc tacgtagata acgaaaagcg caatagccgg
      721 catcccttct ttgcgactat cgaggaggag gtggaatacc ataaaaatta ccccaccatt
      781 taccacttga gagaggaatt ggtcaacagt tctgaaaaag ctgacctgcg gcttgtgtat
      841 ctggcccttg cacacatcat caagtataga ggcaattttc tgatcgaagg cgccctggat
      901 acacaaaaca cttcagtgga tggaatctat aaacagttta tccagacata caaccaggta
      961 tttgctagcg gcatcgagga cggcagcctc aaaaaactcg aggacaataa agacgtggcc
     1021 aaaatcctgg tcgagaaagt tacccgaaag gagaagctgg agcggatcct caagctgtat
     1081 cctggcgaga agtcagctgg catgtttgcc cagttcatca gcctgattgt tggcagcaag
     1141 ggcaattttc aaaagccatt tgaccttatt gaaaagtccg acatagagtg cgccaaggat
     1201 agttacgagg aggacttgga atccctgctc gccctcattg gcgacgaata tgccgagctg
     1261 tttgtggcgg cgaagaacgc ttactcagca gtcgtgctgt cctccatcat tacagtagcc
     1321 gagactgaaa ccaacgccaa gctgtctgca agtatgatag agcgcttcga cacacacgaa
     1381 gaggacttgg gagagctcaa agctttcatc aaattgcacc tgccaaagca ctacgaggaa
     1441 attttttcaa atacagaaaa acacggctac gctggataca ttgacggtaa gactaagcag
     1501 gcagacttct acaaatacat gaaaatgact ttggaaaaca tcgagggcgc tgattacttt
     1561 attgctaaaa ttgaaaagga aaacttcctg cgcaaacaga ggacgtttga taatggagca
     1621 ataccgcatc aattgcatct tgaggagctg gaggctatac tgcaccagca ggccaagtac
     1681 taccccttcc tcaaagagaa ctacgataaa atcaagtcac tcgtaacatt tcggatccca
     1741 tactttgtag ggcctctggc taacggacaa tccgagttcg cctggttgac caggaaagcc
     1801 gacggggaaa tcaggccatg gaacatcgag gagaaggttg attttggcaa gagcgcagtt
     1861 gactttatag agaagatgac aaacaaggac acctatctgc ccaaagaaaa tgtcttgcct
     1921 aagcatagtc tctgctatca aaagtacctg gtgtacaacg aacttactaa ggtgcggtat
     1981 atcaacgacc aggggaagac ctcatatttt tccgggcaag aaaaggagca gatatttaat
     2041 gacttgttta agcagaagag aaaggtcaag aaaaaggatc tggaattgtt cttgagaaac
     2101 atgtcacatg ttgaatcccc caccatcgag ggactcgagg attccttcaa ctccagctac
     2161 agtacgtacc acgacctgct gaaggtgggc atcaaacagg agatcctcga caatcccgtt
     2221 aacactgaga tgcttgaaaa cattgttaag atcctcactg ttttcgaaga taagcgcatg
     2281 atcaaagagc aacttcagca gttcagcgat gtgctggacg gggttgtact caagaaactg
     2341 gaaaggcggc actacaccgg ctggggccgc ctgtccgcca aactgctgat gggcatcagg
     2401 gacaaacagt ctcaccttac aatactcgat tatctgatga atgacgatgg cctcaatcgg
     2461 aatctcatgc agctcatcaa cgattctaat ctctctttca agtccatcat tgagaaggag
     2521 caggtgacca cagccgacaa agacatccag tcaattgtcg ctgacctggc aggctcccca
     2581 gctattaaga aaggcatatt gcagagtctc aaaatagtgg atgaactggt ttctgtgatg
     2641 gggtaccctc cacagaccat cgtggtggaa atggcgcgcg agaaccagac aaccgggaaa
     2701 ggaaaaaaca attccagacc tagatataaa tccctggaga aggccatcaa ggaattcgga
     2761 agccaaattc tcaaggagca ccccacagac aatcaggagc tgaggaacaa tcgcctctac
     2821 ctgtattacc tccagaacgg caaggatatg tacaccggtc aggatctcga tatccataat
     2881 ctcagcaatt acgacattga ccacattgtg ccccagtctt tcattacgga caatagtatc
     2941 gacaacctgg tcctgacctc ttctgctggt aaccgggaaa aaggagacga cgttcctcct
     3001 ctggagatcg tgcggaaacg aaaggtcttt tgggaaaaac tgtaccaggg caacctgatg
     3061 tccaaacgaa aattcgatta cctgacgaag gctgagaggg gtggacttac agaagccgac
     3121 aaagctcggt tcatacatag acagctggta gagacaagac agatcaccaa gaacgtggcg
     3181 aatatcctgc accaacgctt caattacgag aaggatgacc atggtaacac catgaaacag
     3241 gtgcgcatcg tcacactcaa gtctgccttg gtcagccagt ttcgcaagca gttccaactc
     3301 tataaggtga gagatgttaa tgattatcat cacgcccacg atgcatacct gaacggcgtg
     3361 gttgctaata cgctgcttaa agtctatcca cagctcgagc ctgaatttgt gtatggagac
     3421 taccaccagt ttgattggtt taaagccaac aaggccactg caaagaagca attctacacg
     3481 aacatcatgc tgttttttgc ccagaaggat aggatcattg acgagaacgg ggagattctg
     3541 tgggataaaa aatacttgga caccgtcaag aaagtgatga gttatagaca gatgaacatc
     3601 gtcaaaaaga ccgagatcca gaagggcgaa ttctccaagg caaccatcaa acccaagggt
     3661 aattcatcaa agctgattcc ccggaaaaca aactgggatc caatgaaata tgggggcctg
     3721 gatagcccga atatggcgta cgccgtggtg atcgagtacg caaaaggcaa aaacaagttg
     3781 gtgtttgaga aaaaaattat ccgcgtgacc ataatggaaa gaaaagcctt tgagaaagac
     3841 gagaaagcat ttctggagga gcagggttac aggcaaccta aggtattggc taaactcccc
     3901 aaatataccc tgtatgagtg tgaagagggg aggcggagaa tgcttgcgag cgccaacgag
     3961 gcccagaagg gcaaccagca agtgctgcct aatcatttgg tgaccctgct gcatcatgct
     4021 gccaattgtg aagtgagtga cgggaaatca ctggattaca tcgagtcaaa tagagagatg
     4081 tttgccgagc tcctcgccca cgtctctgag ttcgccaagc gctatacact ggccgaagcc
     4141 aatctcaaca agattaacca gctgttcgag cagaataagg aaggtgatat aaaggcaatc
     4201 gcccagtcat ttgtcgacct gatggcattc aacgcaatgg gagcgccggc gagcttcaag
     4261 ttctttgaaa ccacaattga gcggaagcgg tacaacaacc tgaaggaact tttgaattcc
     4321 actattatct accagagtat caccggcctt tacgagagcc gaaagaggct ggatgatagc
     4381 agggctgacc ccaagaagaa gaggaaggtg aggtccatgg tcatagctgt ttcctgtgtg
     4441 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4501 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4561 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4681 tcggctgcgt cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4741 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5041 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5281 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5581 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5641 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5701 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5761 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5821 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5881 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5941 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6001 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6061 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6121 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6181 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6241 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6301 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6361 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6421 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6481 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6541 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6601 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6661 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     6721 ctctgacaca tgcagctccc ggagacggtc aca
//
