LOCUS       sequence-201886-        6114 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 6114)
  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 6114)
  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-Q03LF7
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     311..333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     325..342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     326..342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3760..3780
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3790..3806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3790..3806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3803..3825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3814..3830
                     /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(3838..3844,3845..3862,3863..3868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3883..3904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4242..4830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4322..4341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5001..5792,5793..5861))
                     /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     5624..5643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5862..5966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6034..6052
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6090..6112)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
       61 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      121 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      181 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      241 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      301 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgtgctgtc tcatcatttt
      361 ggcaaagaat tcgcggccgc caccatgtca gatcttgtct tgggcctgga tatcggcata
      421 ggatctgtcg gcgttggcat actcaacaag gtcactggag agattatcca taaaaactct
      481 aggatctttc ctgccgctca ggccgagaac aacttggtgc ggaggactaa ccggcaaggc
      541 agacggcttg cccggcgaaa aaagcaccgc agggtgagac tgaatcgcct tttcgaagag
      601 agtgggctta taaccgactt cactaaaatt agcattaacc ttaacccgta tcagctcaga
      661 gttaaaggac tgacagacga gctctcaaac gaggagcttt ttattgccct gaaaaacatg
      721 gtgaaacaca ggggcatttc ctacttggat gacgccagtg acgacgggaa ctccagcgtg
      781 ggcgactatg ctcagatcgt gaaagagaat agtaagcaac tcgaaactaa gacaccaggc
      841 caaatccagc tggaacgcta tcagacctat ggacagctgc gcggagactt cacagtggag
      901 aaagacggca agaagcatag actcattaat gtgtttccga cgtcagcata ccggtccgaa
      961 gccctgcgaa tattgcagac acaacaggag ttcaatcctc agatcaccga cgagtttatc
     1021 aatcggtacc ttgagatttt gaccgggaag cgcaaatact atcatgggcc aggcaacgaa
     1081 aagtcacgaa ccgactatgg ccgataccga accagtggag aaactctcga caacatcttc
     1141 ggaatcctga taggcaagtg taccttctac ccagacgagt tccgggctgc caaggcaagt
     1201 tacaccgccc aggagttcaa tctgctgaac gacctgaata acctcactgt tcccacagaa
     1261 accaagaagt tgtctaaaga acagaaaaac cagatcatca attacgttaa aaacgaaaag
     1321 gccatgggcc ccgctaaact gtttaagtac atagcgaaat tgctgtcctg tgatgtagct
     1381 gacatcaaag gctatcgcat tgataagtca ggcaaagccg aaatccatac cttcgaggcc
     1441 tacagaaaga tgaagaccct ggagacactt gacatcgaac aaatggacag agagacactg
     1501 gataaacttg cctatgtcct gacactcaat actgagagag agggaatcca agaagccctg
     1561 gaacacgagt ttgcagatgg atctttctca cagaagcagg tggatgagtt ggtgcagttt
     1621 cgcaaggcga attcaagcat ttttgggaag ggctggcata acttctcagt caaactgatg
     1681 atggagctca tcccagagtt gtacgagact tcagaggagc aaatgaccat attgacccgg
     1741 ctgggaaaac agaaaactac ttcttccagc aataagacaa aatatatcga tgagaaactc
     1801 ctgacagaag agatctacaa tcctgtggtg gctaaatccg tgcgccaggc catcaagatc
     1861 gtcaatgcag ccataaagga gtacggcgac ttcgataata ttgtgataga aatggctcgc
     1921 gaaacgaacg aggatgacga aaaaaaagct attcagaaga tacagaaggc taataaggac
     1981 gaaaaagacg cggcgatgct gaaggcggcc aaccagtata acggaaaggc cgagctgccg
     2041 cactccgtct tccacggtca caaacagctt gccacaaaaa tcaggctgtg gcatcagcaa
     2101 ggcgaacgct gcctctacac aggcaaaact atcagtatac acgatctgat caataactcc
     2161 aatcagtttg aggtcgacca catcctgccc ctctccatca cgtttgacga ctcccttgcc
     2221 aacaaagttc ttgtgtacgc caccgctaac caagagaaag ggcagcgcac cccttaccag
     2281 gccctggact ccatggatga tgcctggtca ttccgggaac tgaaagcctt tgtgcgggag
     2341 tcaaagacac tctctaacaa gaagaaggaa tacctgttga ctgaggaaga cattagtaaa
     2401 ttcgatgtcc ggaagaagtt catagaacga aatctggtgg atactcgcta tgcttcacgc
     2461 gtcgttctga acgccctgca ggagcacttc cgagctcata agattgacac caaggtcagc
     2521 gtggtacgag ggcagttcac atctcaactg agaagacact ggggtatcga gaagacgcga
     2581 gatacctatc accaccacgc ggtggacgcc cttataattg cggcctcttc ccagctcaat
     2641 ttgtggaaga agcaaaaaaa taccctcgtc agctacagtg aagatcaact gctggacatc
     2701 gagactgggg aactcatttc cgacgacgaa tataaagaga gtgtcttcaa agccccatac
     2761 cagcatttcg tggatacact gaagtccaag gagttcgagg actccatcct gttcagttac
     2821 caggtggact ccaagttcaa tcggaaaatc tcagatgcaa ccatttatgc cacaaggcag
     2881 gcaaaggtcg ggaaggataa agccgacgag acatatgtgc tcgggaagat taaagacatt
     2941 tacacccaag acgggtacga cgcattcatg aaaatataca aaaaggataa gtcaaaattt
     3001 ctgatgtatc ggcatgatcc acaaacgttc gaaaaggtta ttgaacctat cctcgaaaat
     3061 tatcctaata agcagatcaa tgagaagggg aaggaagtgc cttgtaaccc ttttttgaaa
     3121 tacaaagaag agcacggcta catccgcaag tactctaaaa aaggtaacgg ccctgagata
     3181 aaatcactga agtattacga ctctaaactg gggaaccata ttgacatcac gcctaaagat
     3241 tccaataata aagttgtgct ccagtctgtg tccccgtggc gggccgacgt ctacttcaac
     3301 aagaccactg gaaaatatga gatcctgggg cttaaatacg cagatttgca gttcgaaaaa
     3361 ggaacaggaa cctataagat atctcaggaa aaatacaatg acattaaaaa gaaggagggg
     3421 gtcgactcag actcagaatt taaatttacc ctttacaaaa acgatttgtt gctcgttaaa
     3481 gatacagaaa ccaaggagca gcagctgttc cggtttctgt ccagaacaat gccaaaacag
     3541 aaacactacg tcgagttgaa accttacgac aaacagaaat tcgagggggg ggaggccctc
     3601 atcaaagtcc tcggaaacgt ggccaactcc ggccagtgta aaaaaggact ggggaagagt
     3661 aatatcagca tatataaggt caggactgat gtgctcggca atcagcatat tatcaagaac
     3721 gagggggata aaccaaaact cgactttagc agggctgacc ccaagaagaa gaggaaggtg
     3781 aggtccatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3841 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3901 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3961 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgt cttccgcttc ctcgctcact
     4081 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4321 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4681 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4801 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4921 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4981 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5041 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5101 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5161 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5221 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5281 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5341 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5401 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5461 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5521 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5581 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5641 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5701 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5761 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5821 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6001 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6061 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggag
//
