LOCUS       sequence-201813-        7017 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 7017)
  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 7017)
  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-E7H313
FEATURES             Location/Qualifiers
     source          1..7017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..29)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     129..148
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     342..364
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     356..373
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     357..373
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4694..4714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4724..4740)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4724..4740)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4737..4759)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4748..4764
                     /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(4772..4778,4779..4796,4797..4802))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4817..4838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5176..5764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5256..5275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5935..6726,6727..6795))
                     /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     6558..6577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6796..6900)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6968..6986
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
       61 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      121 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
      181 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
      241 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      301 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      361 aacgacggcc agtgtgctgt ctcatcattt tggcaaagaa ttcgcggccg ccaccatgac
      421 acaaagtgag cggcgcttta gctgtagcat cggaatagat atgggggcta agtataccgg
      481 cgtattctat gcgctctttg accgcgaaga gctccccacc aatctcaact caaaggcaat
      541 gacccttgtt atgcccgaaa ccggcccaag atacgtgcag gcacaacgca ccgccgttcg
      601 gcacaggctg cggggtcaga aacggtatac actggctcgc aaacttgcat tcctcgtagt
      661 ggatgacatg atcaagaagc aggagaagcg cctgacagat gaggagtgga agcgagggcg
      721 cgaagctctc agtgggctgt tgaaaaggag aggttacagt aggccgaatg ccgatggtga
      781 ggatctgacc ccactcgaga acgttagggc agatgtcttc gccgcacatc cagcgtttag
      841 cacttacttt tctgaggtgc ggagcctggc agagcagtgg gaagagttca ccgccaatat
      901 cagtaatgtc gagaagtttc tgggcgatcc caacatcccg gccgacaaag agtttatcga
      961 attcgcagtg gctgaagggc tgatcgataa aaccgagaag aaggcctacc aatctgcact
     1021 gtccaccttg agagctaacg ccaacgtcct gactggcttg aggcaaatgg gacataagcc
     1081 cagaagcgaa tactttaaag cgattgaagc cgatctgaag aaggactctc ggctggccaa
     1141 gattaacgag gcatttggag gagctgaaag gctggctagg ctcttgggaa acctgagtaa
     1201 cctgcagctg agggccgaga gatggtattt taatgctcct gacattatga aggatcgcgg
     1261 ctgggaacca gatcggttta agaagacttt ggtgcgcgcc tttaagtttt tccatccagc
     1321 aaaagaccag aataaacagc acctggaact gattaagcaa atagagaaca gcgaggatat
     1381 catcgaaacc ctttgcaccc tcgatcccaa cagaaccatc cccccatatg aggaccagaa
     1441 caaccggcgg ccccccctcg atcagactct gctcctctct cccgaaaaac ttacccgcca
     1501 gtatggcgaa atttggaaga cctggagcgc ccggcttact tctgctgagc ccaccctggc
     1561 ccccgccgcg gaaattcttg aacggtcaac agacaggaaa tcaagagttg ccgttaacgg
     1621 tcatgagcct ctcccaacgc tcgcatatca actgagttac gcactccagc gagcatttga
     1681 cagatccaaa gccctcgatc cctacgctct gcgggctttg gcggcaggga gcaaatccaa
     1741 caagctgact tctgcccgca ccgccctcga aaattgcatt ggagggcaga atgttaagac
     1801 tttcctggac tgcgcccgcc ggtactatcg cgaggctgac gacgccaagg ttggcttgtg
     1861 gtttgataac gcagacggac tgctggaaag gagtgacctc cacccaccca tgaaaaagaa
     1921 aatacttccg cttctcgtag cgaacatcct gcaaacagac gagacaacag ggcaaaagtt
     1981 cttggatgaa atttggcgca agcagatcaa gggaagagaa accgtcgcgt ccaggtgcgc
     2041 ccggattgaa accgtgcgaa agtcctttgg cgggggattc aacatcgcat acaacacagc
     2101 ccagtatcgc gaagtaaata agcttcctcg gaatgcacag gataaggagc tgctgacaat
     2161 tcgcgatcgg gttgcggaga ctgccgattt tattgcagcg aatctcggtc tttctgacga
     2221 gcaaaagaga aaattcgcta acccctttag tctggcacag ttttataccc tgattgagac
     2281 agaagtcagc gggttctctg cgaccacact ggcagtgcat ctcgaaaacg catggcgcat
     2341 gactataaag gatgcggtaa tcaacgggga aaccgttcga gcagcgcaat gcagcaggct
     2401 gccagccgag acagcacgcc catttgacgg cttggtcagg agactggtcg ataggcaggc
     2461 gtgggaaata gccaagcggg tttccacaga tatccagtct aaggttgact tttctaatgg
     2521 aatcgtagac gttagcatct tcgtggaaga gaataagttt gagttttccg cctccgtcgc
     2581 ggatcttaag aaaaacaagc gggttaagga caagatgttg agcgaggccg agaagctgga
     2641 gacacggtgg ctgataaaga atgagcgcat aaagaaagca agccgcggaa cctgccccta
     2701 caccggcgac cgcttggccg aaggaggaga gatcgaccac attctcccga gatccctgat
     2761 taaagatgca agaggaatcg tgttcaatgc agagcccaac ctgatctatg cctcctccag
     2821 gggcaatcag ttgaagaaga accagcggta tagcctttca gacctgaagg caaattaccg
     2881 gaatgagatc ttcaagacat ccaacattgc cgccatcact gccgagatcg aagacgtggt
     2941 cacaaagctg cagcagaccc accgcctgaa gtttttcgat ctcctgaacg agcatgaaca
     3001 ggattgtgtc aggcacgccc tgtttctgga tgacgggtca gaggctcgcg atgcagtcct
     3061 ggaactgctt gccactcagc ggcgcacgag ggtgaacggt acacagattt ggatgattaa
     3121 aaatcttgct aacaagattc gggaggaact gcagaactgg tgtaaaacaa ccaacaatag
     3181 gctgcacttt caggcagctg cgaccaatgt gtctgacgca aaaaacttga ggctgaagct
     3241 ggcccaaaat cagccagact tcgaaaaacc cgacattcag ccgatcgcct cacattcaat
     3301 tgatgcactg tgctcattcg cagtgggctc agcagatgct gagcgagatc agaacgggtt
     3361 cgattatttg gacggcaaga ccgtgctggg gctctatcct caatcatgtg aggtaataca
     3421 tctccaggcc aagccacaag aggaaaagag ccacttcgat tctgtcgcca tattcaaaga
     3481 aggtatctac gcggaacaat ttttgcctat attcactctt aatgaaaaaa tttggatagg
     3541 ctatgagacg ttgaatgcga agggggagcg gtgtggtgct attgaagtga gtggcaagca
     3601 gcccaaggag ctgctggaga tgctcgcccc atttttcaat aaacctgtcg gtgacctgag
     3661 tgctcacgcc acatatcgaa tcctgaaaaa gcctgcctat gagtttctgg cgaaagcagc
     3721 cttgcagccc ctcagtgccg aagaaaaacg cctcgcagcc cttctcgacg ctctccggta
     3781 ttgtacttct cggaagtcac tgatgtctct gtttatggcc gctaacggaa aatctcttaa
     3841 aaagcgcgag gacgtgctca agcctaaact ctttcaactg aaggtagagc tgaaggggga
     3901 aaaaagcttt aaactgaatg gcagtctgac attgccggtg aagcaagact ggttgcggat
     3961 ctgcgattca ccagaactgg ctgacgcatt cgggaagccc tgtagtgctg acgagttgac
     4021 cagtaagctt gctcgcatct ggaaacgacc agtgatgagg gacttggcgc acgcacctgt
     4081 tcgcagagag ttcagcctgc ctgctatcga taatccctcc gggggatttc gaattaggag
     4141 gacaaatctt tttggtaacg agttgtatca ggtgcacgca attaatgcca agaagtatag
     4201 ggggttcgcc agtgctgggt ccaatgtaga ttggtctaaa ggaattctgt tcaacgaatt
     4261 gcagcatgag aacctcacag aatgcggggg aagattcatc acctctgcag acgtaacccc
     4321 aatgtccgag tggaggaagg tcgtggctga ggataacctt tccatctgga ttgccccagg
     4381 cacagaaggg cggaggtacg tgagggtgga gacaaccttt attcaagctt ctcattggtt
     4441 cgaacagagc gtggagaact gggcaattac ctcccccctg tcactccctg cctccttcaa
     4501 agtagataag cccgccgaat ttcagaaggc tgtgggcacc gagctttcag agctgctggg
     4561 gcaacctcgc agcgagattt ttattgagaa tgtaggaaac gctaaacaca ttcgcttttg
     4621 gtatattgtg gtgtcctcca ataagaagat gaatgagagt tataacaatg tgagtaagtc
     4681 aagcagggct gaccccaaga agaagaggaa ggtgaggtcc atggtcatag ctgtttcctg
     4741 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4801 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4861 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4921 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4981 tcgttcggct gcgtcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5041 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5101 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5161 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5221 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5281 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5341 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5401 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5461 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5521 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5581 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5641 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5701 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5761 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5821 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5881 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5941 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6001 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6061 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6121 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6181 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6241 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6301 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6361 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6421 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6481 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6541 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6601 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6661 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6721 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6781 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6841 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6901 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6961 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacgg
//
