LOCUS       sequence-201809-        7011 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 7011)
  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 7011)
  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-E3ELL7
FEATURES             Location/Qualifiers
     source          1..7011
                     /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             4622..4642
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4652..4668)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4652..4668)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4665..4687)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4676..4692
                     /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(4700..4706,4707..4724,4725..4730))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4745..4766
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5104..5692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5184..5203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5863..6654,6655..6723))
                     /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     6486..6505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6724..6828)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6896..6914
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     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 tgagcagaaa
      361 aaactatgtc gacgactacg caataagcct ggacatcgga aacgcttccg tggggtggag
      421 cgcctttacc cctaactatc gactggtgcg cgccaagggt cacgagctca tcggcgttag
      481 actctttgat ccagccgata ctgcagaaag tagaagaatg gcgaggacaa ccagacgcag
      541 atactctcga aggcgctggc gactgcggtt gttggatgca ctgttcgatc aggctcttag
      601 tgaaattgat ccatccttcc ttgctaggag aaaatactcc tgggtgcatc ctgatgatga
      661 aaataacgcc gattgttggt atggctctgt cctgttcgat tctaacgaac aggataagag
      721 attttatgag aaatatccca caatttacca ccttaggaag gcacttatgg aggacgacag
      781 ccaacatgat atacgggaga tttacctggc catccaccac atggtcaaat atcggggcaa
      841 ctttctggta gaaggcacac tggagtcctc caacgctttt aaggaggacg agctgctgaa
      901 gctgcttgga aggatcaccc gatatgagat gagcgaaggc gagcaaaatt ctgatatcga
      961 gcaggatgac gagaacaaat tggtggcacc cgcaaacggc caattggctg acgccctctg
     1021 tgcaaccaga ggcagccgct caatgcgcgt ggacaacgcg cttgaggcac tgtctgcggt
     1081 gaacgatctc agccgcgagc aaagggcaat cgttaaagcc atctttgctg gcctcgaggg
     1141 taacaagctg gacctggcaa agattttcgt ctcaaaggag tttagttctg aaaacaagaa
     1201 gatcctcggc atctacttca ataagtcaga ctacgaggag aagtgtgtcc agatcgtcga
     1261 ctccggactg ctcgacgacg aggaacgcga attccttgat cggatgcaag gccagtacaa
     1321 cgcaatcgct ttgaagcaac tgctgggcag gtctacaagt gttagcgaca gcaagtgcgc
     1381 atcatacgac gcacacagag cgaactggaa tctcatcaaa ctgcaactca gaaccaaaga
     1441 gaacgagaag gacatcaacg agaactacgg catccttgtc gggtggaaaa ttgactcagg
     1501 gcaaagaaag tccgtgaggg gcgagtccgc ttacgaaaat atgcgcaaga aggctaatgt
     1561 cttttttaaa aagatgattg agacaagcga cctgtcagag acagacaaaa accgactgat
     1621 ccatgatatc gaggaggata agctgttccc gatacagaga gattctgaca atggagtcat
     1681 cccacaccaa cttcaccaaa acgagttgaa gcagatcatt aagaaacagg gaaagtacta
     1741 tccatttctg ctggacgcat tcgagaagga tggtaaacaa attaacaaaa tcgagggcct
     1801 cttgacattc agggtgccat attttgtggg acccctcgtt gtcccagaag acctgcaaaa
     1861 atccgataat agcgagaacc attggatggt aaggaagaag aaaggagaga tcaccccctg
     1921 gaacttcgac gagatggtcg acaaggatgc tagcgggaga aagtttatag agcgattggt
     1981 tggcacggac tcctaccttc tgggagagcc cacactgcct aagaattctc tcctgtatca
     2041 ggagtacgaa gtgttgaacg agctcaataa tgttcgcctg agtgtcagga ccggcaacca
     2101 ctggaatgat aaaaggagaa tgagactggg cagagaggaa aagacgctgt tgtgtcagag
     2161 gctcttcatg aagggacaaa cggtgaccaa gcgcactgcc gagaacctcc tgcgcaagga
     2221 gtacggtcgc acctatgagc ttagcggact gtcagacgaa agtaaattta cctcaagtct
     2281 gagtacatac ggcaaaatgt gcagaatctt tggtgaaaag tacgttaacg agcaccggga
     2341 tttgatggag aagattgttg aactgcagac cgtatttgag gacaaggaaa ccctcctgca
     2401 ccagctgcgc caactggagg ggatatctga ggcagattgt gcactgctgg tgaacaccca
     2461 ctatacaggc tggggccgcc tctcaagaaa gttgctcaca actaaagctg gcgagtgtaa
     2521 gatcagcgat gatttcgcac cccggaaaca ctcaattatt gagattatgc gcgccgagga
     2581 ccgcaatctg atggagataa tcacagataa gcaactggga ttcagtgatt ggatcgagca
     2641 ggagaatctc ggagccgaga acggcagcag ccttatggaa gttgttgatg acctgagagt
     2701 gtctcccaaa gtcaaacgcg gtattattca gagtattcga ctgattgatg acattagtaa
     2761 ggccgttggg aaacgcccgt ctcgaatctt cttggaattg gctgatgaca ttcagccttc
     2821 cggaagaact atctcacgaa aatcccgact ccaagatctc tatcgaaacg ccaacctggg
     2881 taaagagttt aagggtatag cagatgagct taacgcctgt agcgataaag atctgcagga
     2941 tgatcggctg ttcctttatt acacacagct gggaaaagat atgtataccg gagaggagct
     3001 cgacttggac cgattgtcca gcgcttacga catcgatcat attattccac aggcagtcac
     3061 tcaaaacgac tccatcgaca accgggtgct cgtagccaga gcagagaacg cacgaaagac
     3121 cgactccttc acatatatgc cccagattgc agataggatg agaaactttt ggcagatcct
     3181 cctggacaat gggctgatct ctcgcgtgaa attcgagcgg ctcacaagac agaacgaatt
     3241 ctccgaaagg gagaaggaga ggttcgtcca gcggtcactg gtcgaaacca ggcagattat
     3301 gaaaaatgtg gcaacactga tgcgccagag atacgggaat tccgcggccg tgattggact
     3361 gaatgcggag ctgacgaagg agatgcaccg gtacttggga ttcagccaca aaaataggga
     3421 tattaatgac taccaccacg cccaggatgc gctttgcgtg ggaatagccg ggcaatttgc
     3481 ggctaatagg gggttctttg cagacggcga ggtttccgac ggtgcccaga acagctataa
     3541 tcagtacctt agagattacc tgcgaggcta cagagagaaa ctctccgcgg aggatagaaa
     3601 gcaagggaga gcctttgggt tcattgtggg cagcatgcgg agccaagacg agcagaagag
     3661 ggtaaatccc aggacaggag aggtggtctg gagcgaagaa gacaaggact acctcaggaa
     3721 agtgatgaac tatcgaaaga tgctggtcac tcagaaagtg ggagatgatt ttggggctct
     3781 gtacgacgag acaagatacg cagccaccga tccaaagggg attaaaggga ttccttttga
     3841 cggagctaaa caggacacct ccctgtatgg tggattctcc tctgcgaagc ccgcctatgc
     3901 cgtgttgatc gaaagtaaag gcaagacacg actcgtgaac gtgacaatgc aagagtatag
     3961 cctgttgggc gatagacctt ccgatgacga actgcgcaag gtgctggcca aaaagaagag
     4021 cgaatatgcc aaggccaata tacttttgcg ccacgttccc aaaatgcagc tcatcagata
     4081 cggcggggga ctgatggtga tcaagtcagc gggggaactg aacaacgcac agcagctgtg
     4141 gcttccctat gaggaatact gttacttcga tgacctgtca cagggaaagg gcagtcttga
     4201 gaaggatgac ctcaaaaagt tgctcgatag tatcctgggt tctgtccagt gtctctaccc
     4261 atggcacaga tttactgagg aggaactggc cgatctgcac gttgcattcg acaagctgcc
     4321 cgaagatgag aaaaaaaacg ttattaccgg tatagtgtcc gcccttcatg cagacgccaa
     4381 gacagcaaac ctttcaattg tgggaatgac cggcagctgg aggcgaatga acaacaagag
     4441 tgggtacact ttcagcgatg aggacgaatt catctttcaa tccccttctg gattgtttga
     4501 gaagagagtg accgtgggtg agcttaagcg aaaagccaag aaggaggtga actctaagta
     4561 ccgaactaac gaaaagcgct tgccaacact gagtggggct agccagccta gcagggctga
     4621 ccccaagaag aagaggaagg tgaggtccat ggtcatagct gtttcctgtg tgaaattgtt
     4681 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4741 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4801 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4861 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4921 gtcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4981 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5041 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5521 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5641 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5701 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5761 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5821 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5881 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5941 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6001 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6061 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6121 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6181 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6241 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6301 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6361 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6421 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6481 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6541 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6601 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6661 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6721 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6781 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6841 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6901 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6961 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga g
//
