LOCUS       sequence-201893-        6786 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 6786)
  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 6786)
  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-Q8DTE3
FEATURES             Location/Qualifiers
     source          1..6786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     269..291
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     283..300
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     284..300
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4390..4410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4420..4436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4420..4436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4444..4460
                     /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(4468..4474,4475..4492,4493..4498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4513..4534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4872..5460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4952..4971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5631..6422,6423..6491))
                     /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     6254..6273
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6492..6596)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6664..6682
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6720..6742)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
       61 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
      121 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
      181 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
      241 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
      301 gtgctgtctc atcattttgg caaagaattc gcggccgcca ccatgaagaa gccttactcc
      361 atcggcctgg atataggcac taactcagtg gggtgggccg tcgtcaccga cgactataag
      421 gtgcctgcca aaaagatgaa ggttctgggg aataccgaca agtctcatat agaaaaaaac
      481 ttgctgggcg cccttctgtt cgattctggt aataccgctg aggataggag gctgaagaga
      541 accgctcgca ggagatacac tcggaggaga aaccggatcc tttatctgca ggaaatcttc
      601 tccgaggaaa tgggtaaagt agacgacagc ttcttccaca gactggaaga ttcttttttg
      661 gtgactgagg acaagagggg cgaacggcat cctatttttg gtaatctgga agaagaggtg
      721 aagtatcatg aaaacttccc gactatctac catctgcgcc agtacctggc cgacaatccc
      781 gaaaaagtag accttcggct ggtgtacctg gctctggccc atattatcaa gtttcgcggt
      841 cactttctca tcgaagggaa atttgataca cgcaataacg acgtccagcg gcttttccaa
      901 gagttcctgg ccgtctacga caacaccttc gagaacagct ctctgcagga gcagaatgtt
      961 caggtggaag aaatccttac cgataagatc tctaagtcag caaagaaaga ccgggttctg
     1021 aaactgttcc cgaacgagaa gtccaacggt cgatttgcag aatttctcaa actgatcgtg
     1081 ggaaatcaag ccgactttaa gaagcacttt gaactggagg aaaaagctcc actgcaattc
     1141 tctaaggata cttacgaaga agaacttgaa gtgctgctcg cccagatcgg agacaattat
     1201 gccgagctct tcctctcagc caaaaagctg tatgacagta ttttgcttag cgggatactt
     1261 accgtcaccg acgtcggcac taaagcaccc ctcagcgcct ccatgattca gcgatataac
     1321 gaacaccaga tggatttggc acaacttaag cagttcatcc ggcagaagct cagtgacaag
     1381 tataatgagg ttttctccga cgtcagtaaa gatgggtatg ccggctatat tgatggcaag
     1441 acaaatcagg aagccttcta caagtacctg aaaggcttgt tgaacaaaat tgagggcagc
     1501 gggtacttct tggataaaat agaaagggag gactttctta gaaaacagcg caccttcgat
     1561 aatggctcca taccacatca aatccacctt caagaaatga gggccatcat cagacgacag
     1621 gcagaatttt acccgtttct cgctgacaat caggacagga ttgaaaagct gcttaccttt
     1681 cggattccct actacgtggg gccacttgcc aggggtaaat ccgacttcgc ttggctcagc
     1741 cggaagtcag ccgacaaaat caccccttgg aacttcgatg agatcgtgga taaagagtct
     1801 tccgccgagg cgtttatcaa tcggatgaca aactacgact tgtacctgcc gaatcagaag
     1861 gtactgccca agcactccct gctgtacgaa aagtttacag tgtataacga actgaccaag
     1921 gttaagtata aaacagaaca aggtaaaacc gcgtttttcg acgccaatat gaaacaggag
     1981 atcttcgatg gagttttcaa ggtctaccgg aaagtaacca aagataagct catggatttc
     2041 ctggagaagg agttcgatga atttcgaatc gttgacctta ctggcctgga caaagaaaac
     2101 aaggtgttca acgcttctta cggaacctac catgacctgt gcaagatcct cgacaaggac
     2161 tttctggaca acagtaagaa tgagaagatt ctggaagata ttgtgctcac cctgacactg
     2221 ttcgaagacc gggagatgat tcgcaagcgg ctggagaact acagtgacct tctgaccaag
     2281 gagcaggtga aaaaactcga aagaaggcac tatacaggct ggggtaggct ctccgctgaa
     2341 cttatccatg gcatacggaa taaagaatca cgcaaaacta tcctggatta cctgatagac
     2401 gacggtaata gcaatcgaaa cttcatgcag ctcatcaacg acgacgcgct ctctttcaag
     2461 gaggaaatcg ccaaagccca ggtaatagga gaaaccgaca atctcaatca ggtcgtatcc
     2521 gacattgccg ggagccctgc tatcaaaaaa ggaattctgc agagcctgaa gatcgtcgac
     2581 gaactcgtca agatcatggg ccaccagccc gaaaacatcg ttgtagaaat ggcccgcgaa
     2641 aatcagttca ccaaccaggg ccgccgaaat agtcaacaga ggctgaaagg cctgaccgat
     2701 agtattaagg aattcggatc acagatcctg aaggagcacc ctgtcgaaaa tagccagctg
     2761 cagaacgata ggctgtttct ttattatctc caaaacggta gggacatgta cacgggggaa
     2821 gagttggaca tcgactatct ctcacaatat gacatcgacc atatcatccc gcaggcgttc
     2881 atcaaggata atagcatcga caacagagtg ttgacctcta gcaaggagaa tcggggtaag
     2941 tccgacgacg tgccgtctaa ggatgttgtg aggaagatga aatcctactg gagtaaattg
     3001 ctttcagcca agcttatcac ccagaggaaa ttcgacaact tgaccaaagc cgaaagggga
     3061 ggcctgaccg acgacgacaa agcagggttc attaaaagac agcttgtgga gacgagacaa
     3121 atcaccaaac atgtagcgag aatacttgac gaacggttca acaccgagac ggatgagaat
     3181 aataaaaaaa ttcgacaggt caagatagtt acactcaagt caaacctggt ttccaacttc
     3241 agaaaagagt tcgaactgta taaggtcagg gagatcaatg attatcacca tgctcacgac
     3301 gcctatctta atgcagtgat cggcaaagcc ttgctgggag tctatcccca gctcgagccc
     3361 gagtttgtgt atggcgatta ccctcacttc catggccaca aggagaacaa ggctacggct
     3421 aaaaaatttt tctactccaa tatcatgaac ttcttcaaaa aggacgatgt ccgcaccgac
     3481 aagaacggcg aaattatatg gaaaaaagat gagcatatta gtaatatcaa aaaggtgctg
     3541 agctacccac aagtaaatat cgtcaaaaaa gtcgaggagc aaacaggggg tttttctaaa
     3601 gagagtattc tgcctaaagg aaactctgat aagctcatcc ctaggaagac aaaaaagttc
     3661 tattgggata caaagaagta cgggggcttt gattccccta ttgtggctta ttccattctg
     3721 gttattgccg atatcgagaa gggcaagtcc aagaagttga agacagtcaa ggcactggtt
     3781 ggtgttacaa taatggaaaa gatgaccttt gagcgagatc cagtggcctt cctggagcgg
     3841 aagggctacc gcaacgtgca agaggagaat atcattaagc tgcctaaata cagccttttt
     3901 aagttggaaa acgggcgaaa acgccttttg gcttcagccc gggaactgca aaagggaaat
     3961 gagattgtgc tgcctaacca cttgggaact ctgctgtacc acgctaaaaa tatacataag
     4021 gtcgatgagc caaagcacct ggattatgtc gataagcaca aagacgagtt caaggagttg
     4081 ctcgacgtcg tctcaaattt ctccaaaaaa tacacgttgg ctgaggggaa cctggagaaa
     4141 atcaaggagc tgtacgcgca gaacaacggc gaggacctga aagaactggc ttctagcttc
     4201 ataaatctgc tgacatttac tgctattgga gcaccagcaa cgttcaaatt ctttgataaa
     4261 aatatcgacc gaaagaggta cacttccacc accgaaattc tgaacgcaac cctcatccac
     4321 cagtcaatca ctggactgta tgaaacccga atcgacctca acaagttggg cggagacagc
     4381 agggctgacc ccaagaagaa gaggaaggtg aggtccatgg tcatagctgt ttcctgtgtg
     4441 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4501 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4561 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4681 tcggctgcgt cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4741 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5041 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5281 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5581 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5641 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5701 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5761 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5821 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5881 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5941 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6001 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6061 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6121 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6181 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6241 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6301 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6361 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6421 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6481 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6541 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6601 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6661 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     6721 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     6781 agacaa
//
