LOCUS       sequence-201884-        6270 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 6270)
  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 6270)
  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-Q2RX87
FEATURES             Location/Qualifiers
     source          1..6270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     327..349
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     341..358
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     342..358
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3932..3952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3962..3978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3962..3978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3975..3997)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3986..4002
                     /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(4010..4016,4017..4034,4035..4040))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4055..4076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4414..5002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4494..4513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5173..5964,5965..6033))
                     /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     5796..5815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6034..6138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6206..6224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6262..6270,1..14))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
       61 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
      121 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
      181 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
      241 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
      301 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgt
      361 gctgtctcat cattttggca aagaattcgc ggccgccacc atgaggccta ttgagccatg
      421 gattctcggt ctggatatcg gcacagactc tctggggtgg gccgtattca gctgtgaaga
      481 aaaggggccg cctactgcaa aggagctgtt gggcggaggg gtccgactct tcgatagcgg
      541 acgggatgcc aaggatcaca ctagccggca agcagagcgg ggcgccttta ggcgggcgag
      601 acggcagaca agaacctggc catggaggcg cgacagactt attgcgctct tccaggcagc
      661 cggcctcact ccacctgcag ccgaaaccag acaaatagca ttggctctgc ggcgggaagc
      721 agtgtcccgg cctttggctc cagatgccct ctgggccgcc ctcttgcatt tggctcacca
      781 tagggggttt aggagtaata ggattgacaa gcgcgagcgg gcggccgcaa aggcactcgc
      841 caaggcaaag cccgctaaag cgacagcaaa agccacagcc ccggccaagg aagctgacga
      901 tgaggccggc ttctgggaag gagcagaggc cgcacttagg cagagaatgg ctgcaagtgg
      961 tgcaccgact gtgggggccc tgctcgctga cgacctggac agagggcaac ccgtgcgcat
     1021 gcgctacaat cagtcagata gagatggcgt ggtggctccg acacgcgctc tgatcgcaga
     1081 ggaacttgcg gagattgtgg ccagacagtc ctcagcctac ccgggcctgg attggccagc
     1141 ggtgactcgc ctggtacttg accagcgccc cctgcggagc aagggagctg gaccctgcgc
     1201 ttttctgcca ggagaggacc gggccctgcg ggccctcccc acagtgcagg attttataat
     1261 aagacagacg cttgcgaacc tgcgcttgcc aagtacgtcc gccgacgaac cccgcccttt
     1321 gaccgacgag gagcacgcta aggccctcgc actgttgagc actgcccggt tcgtggagtg
     1381 gcctgccctg agacgcgccc tgggcctgaa aagaggagtg aaatttacgg cagaaaccga
     1441 acggaatgga gcaaagcagg cagcgcgcgg cactgcaggg aatctcacag aagccatcct
     1501 ggcacccctg atccccggtt ggagcgggtg ggacctcgac cggaaagaca gggttttcag
     1561 tgacctctgg gccgcacgac aggaccggtc cgcccttctt gctctgattg gagatccccg
     1621 ggggcccact cgcgtgaccg aagatgagac tgccgaggcc gtggccgacg ccatccagat
     1681 agtgttgcct actgggcgag cctcactcag cgctaaggcg gcgcgcgcca tcgcgcaggc
     1741 gatggcccca gggataggtt atgacgaagc ggtcactctt gcgctcggcc tccaccattc
     1801 tcacaggccc cggcaggagc gattggcccg gctgccgtac tacgcagcgg cgctgcccga
     1861 tgtcggtctg gacggtgacc ccgtgggacc accacccgcc gaagatgacg gtgcagcagc
     1921 ggaagcatat tacgggagga taggaaatat atctgttcac atcgcgctga acgaaacacg
     1981 gaagatcgtt aatgcacttc tgcatagaca tggacctatc ctccgcctgg tgatggttga
     2041 aaccaccaga gagctgaagg cgggggccga tgagcgaaag cggatgatag cagagcaggc
     2101 cgaaagggag cgggagaacg ctgagatcga cgtggaactg cggaagtccg accgatggat
     2161 ggctaatgcc cgcgaaagac gacagagagt gcggctggcg cgccgccaaa acaacttgtg
     2221 cccctacaca tctacaccca tcgggcacgc agatttgctg ggcgacgctt acgatatcga
     2281 ccatgtcatt cctctggctc gaggaggccg cgacagtctc gataacatgg ttctctgcca
     2341 atccgatgcc aacaaaacaa aaggagacaa aaccccctgg gaagcatttc acgacaaacc
     2401 tggctggatt gcgcagaggg acgacttttt ggcccgcctg gacccccaaa ctgctaaggc
     2461 cctggcatgg cgatttgcgg atgacgctgg cgagcgcgtg gctaggaagt cagcggagga
     2521 cgaagaccag gggttcctgc ctcggcaact gactgacacc ggatacatcg cgagagtcgc
     2581 cttgcgctat ctctcactgg ttacgaatga accaaacgca gtggttgcaa ctaacggccg
     2641 gctgactgga ttgcttcgcc tggcttggga tatcacgcca gggcccgcac ctagagatct
     2701 gctgcccact cctcgcgatg cactgcgcga cgacactgcg gcaagacggt tcctcgacgg
     2761 cctgaccccg ccaccccttg ccaaagccgt cgagggagcc gtgcaggccc gcctggcagc
     2821 tctggggcga tcacgggtgg cagacgctgg gctggccgac gctctgggcc tgacgctggc
     2881 ttcactcggc ggtggcggga agaatcgcgc agaccaccgc caccacttta tcgatgccgc
     2941 catgatcgcg gtcacaacaa gaggacttat caaccagatt aatcaggctt caggtgcagg
     3001 tcgcattctg gatcttcgga aatggccccg gacaaacttt gaacctccct acccaacgtt
     3061 tagagcagaa gttatgaagc agtgggacca tattcaccct agcattcgcc ctgcacaccg
     3121 cgatggcgga agtctgcatg cagcaactgt ctttggagtg cggaaccggc ccgacgcaag
     3181 ggtgcttgtg cagcgaaaac ccgttgaaaa gttgttcttg gatgccaatg ccaaacctct
     3241 ccccgccgac aaaatcgcag aaattatcga cggcttcgcg tctccccgaa tggccaaaag
     3301 gtttaaggct ctgctggctc ggtaccaggc cgcccacccc gaggtcccac ccgctctggc
     3361 ggctctcgcc gtcgcgagag atcccgcttt tggtcccagg ggaatgactg ccaacacggt
     3421 gatcgcaggc cgaagcgacg gcgatgggga agacgctgga ctgatcactc cgttcagagc
     3481 taaccctaaa gccgcggtgc ggacaatggg gaatgctgtg tacgaggtgt gggagatcca
     3541 ggtgaaaggt cggcctagat ggacgcatag agtcttgact aggttcgaca gaacccaacc
     3601 tgccccccct cctcctcctg agaacgcccg cctcgtcatg cgactgagac ggggagatct
     3661 ggtatactgg cctctggagt ctggagatcg gctgttcctc gtaaagaaaa tggcggtgga
     3721 cgggaggctt gctctttggc ccgcccgctt ggctacagga aaggcaacag ccctgtacgc
     3781 acaactgtca tgccccaata tcaacctcaa cggggatcag gggtactgcg tccagtccgc
     3841 cgaggggatt cggaaggaga aaattcggac cactagctgt accgccctgg gtcgactgcg
     3901 actttcaaag aaggctacga gcagggctga ccccaagaag aagaggaagg tgaggtccat
     3961 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4021 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4081 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4141 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4201 ctgactcgct gcgctcggtc gttcggctgc gtcttccgct tcctcgctca ctgactcgct
     4261 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4321 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4381 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4441 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4501 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4561 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4621 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4681 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4741 acacgactta tcgccactgg cagcaaccac tggtaacagg attagcagag cgaggtatgt
     4801 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4861 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4921 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4981 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5041 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5101 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5161 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5221 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5281 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5341 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5401 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5461 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5521 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5581 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5641 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5701 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5761 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5821 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5881 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5941 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6001 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6061 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6121 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6181 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6241 tttcggtgat gacggtgaaa acctctgaca
//
