LOCUS       sequence-201875-        5937 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 5937)
  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 5937)
  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-I0X7Z6
FEATURES             Location/Qualifiers
     source          1..5937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..136
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     330..352
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     344..361
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     345..361
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3602..3622
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3632..3648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3632..3648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3645..3667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3656..3672
                     /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(3680..3686,3687..3704,3705..3710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3725..3746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4084..4672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4164..4183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4843..5634,5635..5703))
                     /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     5466..5485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5704..5808)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5876..5894
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(5932..5937,1..17))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
       61 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      121 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
      181 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
      241 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      301 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      361 tgtgctgtct catcattttg gcaaagaatt cgcggccgcc accatgatta tgaaactgga
      421 gaagtggagg ctcgggttgg acctgggaac caatagtatc ggatggtctg tgttttcctt
      481 ggataaggat aattccgtgc aggacttgat tgatatgggt gttcgaatat tcagtgatgg
      541 ccgggaccct aagacaaaag aaccgctcgc tgttgctaga cggacggcac gctcccagcg
      601 caaactgatc tacaggcgaa agctgcgccg gaaacaggtt ttcaagttcc tccaggagca
      661 gggactgttc cccaagacta aagaggaatg catgacattg aaatccctga atccgtatga
      721 actgcgcatc aaggccctgg acgaaaaact ggagccgtac gagctcggtc gggcactctt
      781 taatttggct gttagaaggg ggttcaaatc taaccgcaag gacggaagcc gcgaggaggt
      841 aagtgaaaaa aaaagcccgg atgagattaa aacccaggcc gatatgcaga cccatctgga
      901 gaaagcgatc aaggagaacg ggtgccgaac aatcaccgag tttctgtaca agaaccaggg
      961 ggagaacgga ggcatacggt tcgcgccagg caggatgacg tactacccta ccagaaaaat
     1021 gtacgaggag gagtttaatc tcatccggag taaacaagag aaatactacc cccaggtcga
     1081 ctgggatgat atctataagg caatcttcta ccaaaggccc ctgaagcccc agcagagggg
     1141 atactgcatc tatgaaaacg acaaagagcg aacatttaag gccatgccct gcagtcagaa
     1201 gctgaggatc ctccaggaca tcggcaatct cgcttattac gaaggggggt ccaagaagag
     1261 ggttgagctg aacgataacc aagataaagt gttgtacgag ctcctcaata gcaaggataa
     1321 ggtgactttc gaccagatgc gcaaggcctt gtgtctggcg gatagcaact cattcaacct
     1381 ggaggagaac cgcgacttcc tcatcggcaa tcctacagcc gtgaaaatgc gaagcaagaa
     1441 tcgctttggt aagctctggg acgaaattcc tttggaagaa caagatctta taatcgagac
     1501 tataattact gcagatgagg acgacgctgt ctacgaagta ataaaaaagt acgacctcac
     1561 acaggagcag agggatttta tcgtgaaaaa cactatactg cagtccggaa ctagcatgct
     1621 ctgcaaagag gtttctgaaa agttggtgaa aagactggag gagatcgccg acctgaaata
     1681 tcacgaggct gtggagtctt tggggtacaa gttcgccgac cagaccgtcg agaagtacga
     1741 cctgctgcct tactatggaa aggtgctccc cggctccaca atggagattg atcttagtgc
     1801 ccctgaaacg aatcccgaga agcactatgg caaaatatca aatccaaccg tgcacgtggc
     1861 gctcaatcaa acacgagtcg ttgtgaacgc tcttatcaag gagtacggaa aacccagcca
     1921 gattgcaatt gagctgagtc gggaccttaa gaataacgtc gagaagaaag ctgaaatcgc
     1981 caggaagcag aaccagagag ccaaggagaa catcgccatc aatgatacca tttccgccct
     2041 gtaccacaca gctttcccag ggaaaagttt ttacccaaac cggaacgatc gaatgaagta
     2101 ccggctttgg tcagaactgg gcctcgggaa caagtgtatt tattgtggga aaggcatttc
     2161 cggagctgag ctgtttacta aagagattga gattgaacac attctgccgt tctcccggac
     2221 tctgctggat gctgagagta atctcacagt ggctcactcc agctgcaatg ctttcaaggc
     2281 tgagcgatcc cccttcgagg cgttcggaac taaccccagc ggctactcat ggcaagagat
     2341 catacagcgc gctaaccagc tgaaaaacac ctccaaaaag aacaagttta gcccaaatgc
     2401 catggatagt tttgaaaagg actcaagctt tatcgcccgc cagttgagtg acaatcagta
     2461 tatagcaaag gccgctctgc gatacctcaa gtgcctcgtg gaaaacccat ccgacgtgtg
     2521 gacaaccaac ggttccatga ccaaacttct tagagataag tgggagatgg acagcattct
     2581 ttgccggaaa ttcacggaga aggaagttgc cctcctgggc ctgaaacccg aacaaatagg
     2641 gaattataag aaaaacaggt ttgatcatcg gcatcacgcc attgatgccg tagttattgg
     2701 actgacagac agatccatgg tgcaaaagct ggcaaccaag aacagccata agggcaacag
     2761 aatcgagatc ccggagtttc cgatcctgcg gtctgacctc attgagaagg ttaagaacat
     2821 tgtagtatcc tttaaacccg atcacggggc tgagggcaag ctgtctaaag aaacactgct
     2881 gggaaagata aagctccacg ggaaagaaac tttcgtttgt cgcgaaaata tcgtgtcttt
     2941 gtccgagaag aatcttgatg acatcgttga tgagaagatt aaatctaagg tcaaagacta
     3001 cgtggccaag cacaagggac aaaagataga agctgtgctg agtgatttct caaaggaaaa
     3061 cggaattaag aaagtaagat gtgtaaatcg ggtacagacg ccaattgaaa ttacctcagg
     3121 taaaatctct cgctatctgt cccccgagga ctacttcgca gcagtcattt gggagatccc
     3181 aggggaaaaa aaaaccttca aagcacagta cattcggaga aacgaggtgg agaagaattc
     3241 taagggattg aatgtggtca aaccagccgt cctcgagaat gggaagccac atcccgccgc
     3301 taagcaagtg tgtctgctcc acaaagacga ttacctggag ttcagcgata aaggcaagat
     3361 gtacttctgc aggatcgccg gatatgcagc cactaataac aagttggata ttagacccgt
     3421 gtacgcagtt agctattgtg cagattggat aaattctact aacgaaacca tgttgaccgg
     3481 gtactggaag cctacaccca ctcagaattg ggtgtccgtc aacgttctgt tcgacaagca
     3541 gaaggcacga ctcgtgactg tatctcctat cggtcgggtc tttcgaaaga gcagggctga
     3601 ccccaagaag aagaggaagg tgaggtccat ggtcatagct gtttcctgtg tgaaattgtt
     3661 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3721 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3781 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3841 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 gtcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4261 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4501 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4861 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4921 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4981 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5041 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5101 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5161 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5221 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5281 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5341 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5401 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5461 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5521 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5581 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5641 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5701 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5761 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5821 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5881 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctg
//
