LOCUS       sequence-201885-        6915 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 6915)
  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 6915)
  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-Q03JI6
FEATURES             Location/Qualifiers
     source          1..6915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     276..298
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     290..307
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     291..307
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4526..4546
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4556..4572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4556..4572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4569..4591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4580..4596
                     /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(4604..4610,4611..4628,4629..4634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4649..4670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5008..5596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5088..5107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5767..6558,6559..6627))
                     /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     6390..6409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6628..6732)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6800..6818
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6856..6878)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
       61 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
      121 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
      181 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      241 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      301 ggccagtgtg ctgtctcatc attttggcaa agaattcgcg gccgccacca tgaccaagcc
      361 atactctatt ggcttggaca ttgggacaaa ttcagtgggt tgggcggtga ctactgacaa
      421 ttacaaggtc ccctcaaaga agatgaaagt gctcggaaac acgagcaaaa agtacataaa
      481 gaagaacctc ttgggggtgc ttctgttcga ttcaggaata accgctgagg ggcgcaggtt
      541 gaaacgcact gccaggcggc gctacactcg gaggcggaac cggattctgt atctgcagga
      601 gatcttcagt acagaaatgg ctactctgga tgatgccttc tttcagcggc tcgatgacag
      661 tttcctggtc cctgacgaca aaagggacag caagtacccc attttcggga acctcgtcga
      721 ggagaaggcg taccacgacg aatttcccac catctatcat cttagaaagt acctggccga
      781 tagcacaaag aaggccgatc tcagacttgt atatctggcc ttggctcaca tgattaaata
      841 ccgcggtcat ttcctgatcg aaggcgaatt caacagtaag aacaacgata tccagaaaaa
      901 cttccaggat tttctggaca cttataacgc catattcgag tcagacctgt cccttgaaaa
      961 tagcaaacaa ctggaggaaa tcgtgaaaga taagattagc aagctggaaa aaaaggatag
     1021 aattctgaaa cttttccctg gcgagaagaa ttccgggatt ttttccgaat ttctgaagct
     1081 gatagttggc aatcaggctg acttccggaa atgtttcaac ctggatgaga aggcgagtct
     1141 gcacttcagc aaggagtctt acgacgaaga cctggaaaca ctgctcggct atatcgggga
     1201 tgattactca gacgtgttcc tgaaagcaaa aaagctttat gacgcaatct tgttgtcagg
     1261 cttcctgact gttactgaca acgagacgga agcacccctc tctagcgcaa tgataaagag
     1321 gtataacgag cataaagagg acttggccct tttgaaggag tacattagga acatcagtct
     1381 caagacctat aacgaggtgt tcaaagacga tactaagaat ggctacgctg gttatattga
     1441 tgggaagacc aatcaggagg atttttatgt atatctcaaa aaactcctcg cagagttcga
     1501 gggagctgat tacttcttgg agaagatcga tcgcgaggac tttctcagga agcagagaac
     1561 cttcgacaat ggttccatcc cttatcagat ccatcttcag gagatgcgcg ccatcctgga
     1621 caaacaggcc aagttctacc ctttcttggc aaaaaataag gaacgcattg agaaaatcct
     1681 gacatttaga atcccttatt acgtgggccc cctggcacgc ggaaattctg acttcgcctg
     1741 gagcattcga aaacggaatg aaaagatcac accttggaac ttcgaggacg ttatagacaa
     1801 ggagtcctcc gctgaagcat tcatcaatcg gatgacctcc ttcgacttgt atttgccaga
     1861 ggagaaggtg ttgccaaaac acagtctgct ctacgagact ttcaatgtgt acaatgagct
     1921 gaccaaagtc agattcattg ccgagtcaat gagagattat cagttcctgg attctaaaca
     1981 gaaaaaagat attgtgcgcc tgtatttcaa ggataaacgg aaagtgacgg acaaagacat
     2041 tattgagtat ctgcacgcca tctacggtta tgacggcatc gaactcaaag gaatcgagaa
     2101 acagttcaac agttccctga gtacttatca cgatctcctg aatatcatca acgacaaaga
     2161 atttcttgac gacagttcaa acgaggctat tattgaagag atcatccaca cactcacgat
     2221 attcgaagac agagaaatga ttaagcagag actgtcaaaa ttcgaaaata tttttgataa
     2281 gtctgtgctg aagaagctca gtaggaggca ttacaccggg tggggcaaac tttcagctaa
     2341 gctgattaac gggatccgag acgaaaagtc agggaataca atcctggact acctgatcga
     2401 cgatgggatt tctaaccgga actttatgca actgatacac gacgatgcat tgtcctttaa
     2461 aaagaaaatc cagaaagccc aaattatagg cgacgaagac aagggcaaca ttaaagaagt
     2521 ggttaaatca ttgcctggga gccccgcgat caagaagggg atactccaat ccatcaaaat
     2581 tgtggatgag cttgtcaagg tgatgggagg ccgaaagccg gaaagcattg tggtggagat
     2641 ggctagagaa aatcagtaca ccaaccaggg gaagagtaat tcccagcagc ggcttaaacg
     2701 gttggaaaag agcttgaagg aactggggtc aaaaattctg aaggagaaca tccctgctaa
     2761 actcagcaag attgataata acgccctgca gaacgatagg ttgtacctgt actacctcca
     2821 gaatgggaag gacatgtaca ctggcgacga tctggatatc gacagactgt ctaactatga
     2881 tatcgatcat attatcccac aggccttcct taaggataat tccatcgata ataaggtgct
     2941 cgtcagctct gcatccaatc ggggtaaatc tgacgacgtg cctagtctgg aggtcgttaa
     3001 gaaacgcaaa acattttggt accaactgct taagtcaaaa cttatcagcc agcggaagtt
     3061 cgataatttg acaaaggccg aacggggcgg cctcagccct gaggataaag ccggtttcat
     3121 ccagaggcaa ctggtggaga ctcggcaaat tacaaaacat gtagctaggc tcttggatga
     3181 gaaattcaac aacaagaagg atgagaataa ccgagccgtc cgaaccgtga agatcattac
     3241 tttgaagagt acgctggtga gtcagtttcg gaaggacttc gagctgtaca aagtgcgaga
     3301 aataaacgat tttcaccatg cccacgatgc ctatctgaac gccgtggtgg cctctgctct
     3361 cttgaagaag tatcccaaac ttgagcccga gtttgtgtac ggtgattacc ccaaatacaa
     3421 ctcctttcga gaaagaaaga gtgctaccga aaaagtctac ttctattcca acatcatgaa
     3481 tattttcaaa aaaagcatct ccctggctga tggtcgagtc atagagcggc ccctcatcga
     3541 agtcaatgag gagacgggcg agagtgtgtg gaacaaggaa tctgatctgg ccacggttag
     3601 aagagtgctt agctacccgc aggtcaacgt tgtgaagaag gttgaagaac aaaatcacgg
     3661 tctggaccgg ggaaagccta aaggcctgtt caatgctaac ctgtcatcaa aaccaaagcc
     3721 taacagcaac gaaaaccttg tcggagctaa ggaatacctg gacccgaaga agtacggtgg
     3781 atacgccggc atctctaatt cctttaccgt gcttgtcaag ggaaccatcg agaagggcgc
     3841 caagaagaag atcacaaacg tcctggaatt tcaggggatc agcatcctgg atcggataaa
     3901 ttaccgcaaa gacaagctta acttcctcct ggagaaaggc tacaaggaca tcgagctcat
     3961 tatcgaactg ccaaaatata gtctgtttga gctttccgac ggtagccggc ggatgctggc
     4021 aagcattctt tcaaccaaca ataagcgggg ggaaatccat aaagggaacc agatcttcct
     4081 tagtcagaag ttcgtcaagc tcctgtacca tgcaaagcgg atatcaaata caatcaatga
     4141 gaatcaccga aagtacgtcg agaatcataa aaaggagttc gaggaactct tttattacat
     4201 cctggaattc aatgaaaact atgtgggagc caagaaaaac ggcaaactgc tgaactcagc
     4261 tttccagagc tggcaaaacc attccatcga tgagttgtgt agttctttta tcggccctac
     4321 cggttccgaa aggaaaggat tgttcgaact cacaagcagg ggttctgcag ctgactttga
     4381 gtttctggga gtgaagatcc caagataccg agattataca ccctctagtc tgcttaagga
     4441 tgccaccctg atccaccagt ctgtgaccgg cctttatgag acaaggatcg acctggctaa
     4501 acttggggaa ggcagcaggg ctgaccccaa gaagaagagg aaggtgaggt ccatggtcat
     4561 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4621 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4681 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4801 cgctgcgctc ggtcgttcgg ctgcgtcttc cgcttcctcg ctcactgact cgctgcgctc
     4861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6721 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6781 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6841 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6901 agcggatgcc gggag
//
