LOCUS       sequence-201892-        6459 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 6459)
  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 6459)
  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-Q6KIQ7
FEATURES             Location/Qualifiers
     source          1..6459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     275..297
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     289..306
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     290..306
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4069..4089
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4099..4115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4099..4115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4112..4134)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4123..4139
                     /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(4147..4153,4154..4171,4172..4177))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4192..4213
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4551..5139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4631..4650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5310..6101,6102..6170))
                     /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     5933..5952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6171..6275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6343..6361
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6399..6421)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
       61 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
      121 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
      181 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
      241 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
      301 gccagtgtgc tgtctcatca ttttggcaaa gaattcgcgg ccgccaccat gtatttctac
      361 aagaataaag agaacaagct gaacaaaaag gtggtgttgg gactcgacct ggggattgcc
      421 tcagtggggt ggtgtctgac tgacatctct caaaaggagg ataacaagtt tcctatcatc
      481 ctccatggag tccgcctgtt cgagacggtg gacgactccg acgataagct gctgaatgaa
      541 acacgaagga agaagagagg acagagaaga cgaaatcggc ggcttttcac ccgcaagcgg
      601 gattttatca agtacctgat cgacaataat attattgaac ttgagttcga caagaatccg
      661 aagattctgg tgcgcaattt catcgaaaag tatatcaatc cctttagtaa gaatctcgaa
      721 ctcaaatata agagcgttac aaatctcccc atcgggttcc ataacctgcg caaggccgca
      781 atcaatgaga aatacaagct tgacaagagt gagctgatcg tgttgttgta tttttacctg
      841 tcccttcgcg gtgccttctt cgataatccc gaagacacga agtctaaaga aatgaacaag
      901 aatgagatcg aaatttttga taagaatgag tccattaaga acgcggaatt cccgatcgat
      961 aagataatcg aattttacaa gatcagtgga aagataaggt ccacgataaa ccttaaattc
     1021 ggccaccagg actacctgaa agaaataaaa caagtgtttg aaaagcagaa catcgatttt
     1081 atgaattacg agaaatttgc aatggaggaa aaaagcttct tttcccggat aaggaattac
     1141 agcgaaggac ccggaaatga gaagtctttc tccaagtacg gattgtatgc aaatgagaac
     1201 ggtaaccccg agttgatcat aaacgaaaag ggacagaaga tttatacaaa aatttttaaa
     1261 acactgtggg aaagtaaaat cggcaagtgt agctacgata aaaagcttta ccgggctccc
     1321 aagaactcct tctcagcaaa agtattcgat atcactaaca aactgactga ctggaaacac
     1381 aaaaatgaat atattagtga gaggttgaaa agaaagattt tgcttagcag attcctgaac
     1441 aaggatagca aatccgctgt ggagaaaata cttaaggagg agaacattaa gttcgagaac
     1501 ctttctgaaa tcgcatataa taaggatgat aataagatca atcttcccat cattaatgca
     1561 taccacagcc tgacaactat ctttaaaaag cacttgatca actttgagaa ttacttgatt
     1621 tctaacgaga atgatctgtc taagctgatg agcttttata agcaacaaag cgaaaaactg
     1681 ttcgtgccta acgagaaagg gtcttacgag atcaatcaaa ataacaacgt actgcacatc
     1741 ttcgatgcca tttccaatat ccttaacaag ttcagtacca tccaggatcg aattcggatc
     1801 ctcgaagggt actttgaatt cagtaatctt aagaaggacg tgaagagctc tgaaatctat
     1861 tcagagattg ctaaactgcg agaattctca ggaaccagct ccctgtcttt tggggcttac
     1921 tacaaattta tccccaacct catctcagag ggctccaaaa actacagcac cataagttat
     1981 gaggagaaag ccctccagaa tcagaaaaat aacttcagtc actctaacct gttcgaaaag
     2041 acttgggtgg aggatctcat tgcctcccct accgtcaagc ggtctttgcg acagactatg
     2101 aacctgctga aggaaatctt taagtattcc gaaaaaaata acctcgagat cgagaagatt
     2161 gtagtcgagg taactagaag ctctaacaac aagcacgaaa ggaaaaaaat tgaaggtata
     2221 aacaagtatc gaaaagagaa gtacgaggaa ctgaaaaaag tatatgatct ccccaacgaa
     2281 aacaccactc tgcttaagaa gttgtggttg ttgcgccagc agcagggcta tgacgcatac
     2341 tcactccgga agattgaagc caatgatgtg attaataagc cttggaatta cgatattgat
     2401 cacattgtgc ctaggtctat ctccttcgac gattcattta gcaatctggt aattgtgaat
     2461 aaactcgata atgcaaaaaa atcaaatgac ctgtccgcca aacagtttat tgagaaaatc
     2521 tatgggatcg agaagctgaa ggaggccaag gaaaactggg ggaattggta cctgcgcaat
     2581 gctaatggta aggctttcaa cgataaaggt aagttcatca aactttatac cattgacaat
     2641 ctcgacgaat ttgataacag cgattttatt aacagaaacc ttagtgacac cagctacatt
     2701 actaacgctc tcgtcaacca tcttaccttt tctaattcta agtataagta ttctgttgtc
     2761 tcagtgaacg gaaaacagac ctccaatctt cgcaatcaga ttgctttcgt cgggattaag
     2821 aacaacaagg aaaccgaacg cgagtggaag cgtccggagg gctttaagtc aattaattct
     2881 aacgatttcc tgatacgcga ggaaggaaaa aacgatgtta aagatgatgt tctcattaag
     2941 gataggtcct ttaacgggca ccatgccgag gacgcctact ttatcacaat catctcccag
     3001 tacttcaggt cctttaagcg gatcgagaga cttaacgtca actacaggaa agagacacgc
     3061 gaactggacg acctcgaaaa gaataatatc aagtttaaag aaaaggcctc cttcgataat
     3121 tttctgttga ttaatgccct ggacgaactg aatgaaaaac tcaatcaaat gcggttcagc
     3181 aggatggtga tcacaaagaa gaatacccag ctgtttaatg aaacactcta tagtgggaag
     3241 tacgacaagg gcaagaacac aatcaagaaa gtcgagaagc ttaacttgct ggacaatcgc
     3301 acagacaaaa tcaagaaaat cgaagagttc tttgacgaag ataagctcaa ggaaaacgaa
     3361 ttgactaaac tccatatatt taaccacgac aagaatctgt acgaaaccct taaaatcata
     3421 tggaacgagg tgaaaatcga aatcaagaat aagaatctta acgagaaaaa ttactttaag
     3481 tacttcgtga ataagaagct gcaggagggc aagattagtt tcaacgaatg ggtgccgatc
     3541 ctggacaacg acttcaaaat tattcgaaaa attcggtata tcaagttctc tagcgaggag
     3601 aaagagactg acgagatcat tttttcccag tcaaacttcc ttaaaatcga tcaaaggcag
     3661 aactttagct tccataatac cttgtactgg gtgcagattt gggtctacaa aaaccagaaa
     3721 gatcagtatt gctttatttc cattgacgcc aggaatagca agtttgagaa agatgaaatc
     3781 aaaatcaact atgagaagct caagacacaa aaggagaaac tgcagatcat aaatgaggaa
     3841 ccgattctca agataaacaa aggtgatctg tttgagaacg aagaaaagga actgttctat
     3901 atcgtgggca gagacgaaaa accacagaag ctcgaaataa aatatatcct ggggaagaaa
     3961 atcaaagacc aaaagcagat tcaaaaacct gtcaaaaaat actttcccaa ttggaaaaag
     4021 gttaatctta catacatggg ggaaatcttc aagaaaagca gggctgaccc caagaagaag
     4081 aggaaggtga ggtccatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4141 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4201 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4261 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcgtc ttccgcttcc
     4381 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4441 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4501 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4741 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4921 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4981 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5041 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5101 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5161 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5221 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5281 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5341 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5401 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5461 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5521 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5581 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5641 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5701 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5761 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5821 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5881 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5941 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6001 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6061 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6121 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6181 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6241 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6301 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6361 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6421 gagacggtca cagcttgtct gtaagcggat gccgggagc
//
