LOCUS       sequence-201878-        6342 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 6342)
  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 6342)
  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-J5TNY4
FEATURES             Location/Qualifiers
     source          1..6342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..88
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     282..304
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     296..313
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     297..313
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3959..3979
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3989..4005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3989..4005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4002..4024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4013..4029
                     /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(4037..4043,4044..4061,4062..4067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4082..4103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4441..5029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4521..4540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5200..5991,5992..6060))
                     /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     5823..5842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6061..6165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6233..6251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6289..6311)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
       61 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
      121 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
      181 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      241 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      301 aacgacggcc agtgtgctgt ctcatcattt tggcaaagaa ttcgcggccg ccaccatgct
      361 catgtccaag cacgtgctgg ggttggatct gggggtcggg tcaattggct ggtgcttgat
      421 tgccctggac gctcagggtg atcccgctga aattctgggc atggggagcc gcgtcgtgcc
      481 cctgaataat gctacaaaag caattgaggc tttcaatgcc ggagctgcct tcactgcaag
      541 ccaagagcga acagcaagaa gaacgatgag acgcgggttc gcccgctatc agttgcgcag
      601 ataccgcctg agaagggaac ttgagaaagt tgggatgttg cccgatgctg ctctgatcca
      661 actgcccctc ctggaactgt gggagcttcg ggaaagagcc gcgaccgcag gccgacggct
      721 gaccctccct gagctgggac gcgtgctgtg ccacatcaat cagaagcgcg gctacagaca
      781 tgtaaaaagc gatgctgctg ccattgtggg cgacgagggc gagaaaaaga aggactctaa
      841 ttctgcgtat ctggccggaa ttagggcaaa tgatgagaag ctccaagctg aacacaagac
      901 tgtcggacag tacttcgccg aacagctgcg gcaaaatcag agtgaaagtc ctactggagg
      961 cataagctac agaattaagg accagatctt ctctcgccaa tgttacatcg acgagtacga
     1021 ccagatcatg gctgtgcaga gagtgcatta tcccgacatt ctgacggacg agtttatcag
     1081 aatgctgagg gacgaggtaa tcttcatgca acgccccctc aaatcttgta agcacctggt
     1141 gtctctgtgc gaatttgaga agcaagagcg cgtcatgcgg gtccagcagg acgacggcaa
     1201 gggaggctgg cagctggtgg aacggagggt caaatttggg cccaaagtgg ccccgaaatc
     1261 ctcaccactg tttcaactgt gctgtatcta cgaagccgtg aataacatcc ggctcacacg
     1321 cccaaacgga tccccatgtg acattacacc tgaggaacgg gctaagatcg tggcccatct
     1381 ccagagtagc gcttctcttt cctttgccgc tttgaagaag ctgctcaaag aaaaagcgct
     1441 gattgcagac cagctcacaa gcaagtccgg tctgaagggt aactccactc gcgtggcact
     1501 tgcttctgct cttcagccct acccccaata ccatcacctg cttgacatgg agcttgaaac
     1561 cagaatgatg accgtccagt tgaccgacga ggaaaccggg gaagttaccg agcgcgaggt
     1621 cgccgtcgta actgactcat acgtgcgaaa gcctctttat cgcctgtggc atatactgta
     1681 cagcatcgag gagcgggagg ccatgcggag agcgttgatc acacagctgg ggatgaaaga
     1741 ggaggatctg gacggaggcc tgctcgacca gctgtaccgg ctggacttcg tcaagcctgg
     1801 ctacgggaac aagtctgcta agttcatttg taagctgctc cctcagctcc agcagggact
     1861 gggctactct gaagcatgcg ccgctgtcgg gtatagacat tccaatagtc caactagtga
     1921 ggagatcacc gagcggactc tgctggagaa aatccctctg ctgcagcgga acgaactgcg
     1981 ccaacccttg gttgagaaaa ttctgaatca gatgatcaat ttggtcaacg ctctgaaggc
     2041 agagtatggt atcgacgagg tccgcgtgga gctggcaaga gaactgaaaa tgtcccggga
     2101 ggaacgcgag agaatggcac ggaacaataa ggaccgggag gagcgaaata aaggcgtggc
     2161 tgccaagatc cgggagtgtg gtctttaccc cacaaagccc agaatccaga aatacatgct
     2221 gtggaaggaa gccggtaggc agtgtctcta ctgcggccga tcaattgaag aggagcagtg
     2281 tcttcgagag ggtggaatgg aggtagagca tattattcct aagtcagtcc tgtatgacga
     2341 ttcatacgga aataagacct gtgcatgcag gagatgcaac aaggagaagg gcaaccggac
     2401 cgctctggaa tacattagag ctaaggggcg cgaggccgag tatatgaaac gcattaacga
     2461 cctgctcaaa gagaagaaga tctcatacag taagcaccag agacttcgct ggttgaagga
     2521 ggacatcccc agcgacttcc tcgagcgaca actgcggctt acccagtata taagtcgcca
     2581 ggcaatggcc attttgcaac agggaatccg cagagtgagt gcttccgaag ggggagtcac
     2641 agctagattg aggtccctgt gggggtatgg caagattctc cacacactca accttgatag
     2701 gtatgatagt atgggcgaaa cagaacgcgt ttctcgagag ggcgaggcca ccgaggaact
     2761 tcatatcacg aactggtcca agagaatgga tcaccggcat catgcaatcg acgctctcgt
     2821 cgtcgcctgt acccgccagt cttatattca gcggctcaat cggcttagta gtgagttcgg
     2881 cagggaggat aagaagaaag aagatcagga ggctcaagaa caacaggcca ccgaaaccgg
     2941 ccggcttagt aacctcgaac gctggcttac ccagcgtccc cacttttctg tgagaacggt
     3001 gtctgacaag gtcgcggaaa ttttgattag ttaccgacca ggccagcggg tagtgactag
     3061 agggagaaac atctaccgca agaagatggc tgacgggagg gaagtgagct gcgtccaaag
     3121 gggtgtcttg gtacctcggg gagagctcat ggaggcgagc ttttatggga agattctctc
     3181 ccaggggagg gtgcgcatcg tgaagagata cccactccat gacctgaaag gtgaggtagt
     3241 ggatccacat ctgcgcgaac tgatcaccac atataatcag gaactcaaaa gccgagaaaa
     3301 aggggctccc atcccccctc tgtgtctgga caaagataag aagcaagagg tccgaagcgt
     3361 gaggtgttac gctaaaactc tgagcttgga caaggcaatt cccatgtgct ttgacgaaaa
     3421 aggcgaaccg actgcgttcg tcaaaagtgc cagtaatcac cacctcgctt tgtatcggac
     3481 acccaagggg aaactggtgg aatccatcgt cacattttgg gatgcggtcg accgagctag
     3541 gtacgggatc cctctggtga ttacccaccc aagagaggtc atggaacaag tgcttcagcg
     3601 cggggacata cctgaacagg tattgagtct gcttccccct tccgactggg tttttgtgga
     3661 ttcactccag caggacgaaa tggtggtcat cggcttgagc gacgaggaac tgcagcgggc
     3721 tttggaagcg caaaattacc gcaaaatctc cgagcacctc tatcgggtgc agaaaatgtc
     3781 ctcaagttac tatgtgtttc gatatcacct tgagacatca gtggctgatg acaagaatac
     3841 ttccggacga atcccaaagt tccaccgcgt tcagtctctg aaagcctacg aggaacgaaa
     3901 tattaggaaa gtccgggtcg acctgctggg acgaatctcc ctgttgagca gggctgaccc
     3961 caagaagaag aggaaggtga ggtccatggt catagctgtt tcctgtgtga aattgttatc
     4021 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4081 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4141 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcgtc
     4261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6181 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6241 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6301 gcagctcccg gagacggtca cagcttgtct gtaagcggat gc
//
