LOCUS       sequence-201817-        7014 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 7014)
  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 7014)
  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-F3UXG6
FEATURES             Location/Qualifiers
     source          1..7014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     326..348
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     340..357
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     341..357
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4675..4695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4705..4721)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4705..4721)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4718..4740)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4729..4745
                     /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(4753..4759,4760..4777,4778..4783))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4798..4819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5157..5745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5237..5256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5916..6707,6708..6776))
                     /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     6539..6558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6777..6881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6949..6967
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(7005..7014,1..13))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
       61 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
      121 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
      181 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      241 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      301 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgtg
      361 ctgtctcatc attttggcaa agaattcgcg gccgccacca tgacgaagtt caacaaaaat
      421 tacagcatcg ggctggacat tggagtgagt tccgtcggat atgccgtagt cactgaggat
      481 tatcgcgtgc ctgcgttcaa gtttaaagtg ctcggcaata cagagaagga aaaaattaag
      541 aagaacctga ttggttccac taccttcgtc agcgcacagc cagccaaagg aactcgcgtg
      601 ttccgcgtta accgccgcag gatcgaccgc cggaatcatc gaatcaccta cctgagggat
      661 atctttcaaa aagaaattga gaaggtcgac aagaacttct atcgcagact ggacgagagc
      721 ttccgggtgc tcggagataa aagcgaagac ctgcagatca aacaaccgtt ctttggtgac
      781 aaggagctcg aaaccgccta ccacaaaaag taccctacga tctaccacct gaggaaacac
      841 ctggctgatg ctgacaagaa tagtccggtt gccgatatcc gggaggtgta catggcaatt
      901 agccatatcc ttaagtaccg ggggcatttc ctgacgctcg acaagatcaa ccctaacaat
      961 attaacatgc agaatagctg gatcgatttt attgagagct gtcaggaggt gttcgacctc
     1021 gaaatcagcg acgagtccaa gaatatagca gacattttta agtcaagtga aaatagacag
     1081 gagaaggtaa agaagatact gccttacttc cagcaagagt tgttgaaaaa agacaagagt
     1141 atttttaaac agttgttgca gctccttttc ggactgaaga ctaagtttaa agactgcttt
     1201 gaactcgaag aggagcctga cctgaatttc tccaaagaaa attatgatga gaacctggag
     1261 aacttcctgg ggtcacttga agaggatttc tcagatgtct ttgcaaagct gaaagtcctc
     1321 agagatacca tactcctgtc tgggatgctc acctacacag gcgccaccca cgcacgcttc
     1381 agtgccacga tggtagagag atacgaagag cacagaaaag atttgcagcg gttcaagttt
     1441 ttcatcaagc agaatctgtc agagcaggat tacttggata tctttggcag aaagacgcag
     1501 aatggatttg atgtggacaa ggagacaaaa ggctatgtgg gctacatcac aaacaagatg
     1561 gtcctgacta accctcagaa acagaagacc attcagcaga acttctacga ttacatctcc
     1621 ggcaaaataa cagggattga gggggcggaa tactttctga acaaaattag cgacgggacc
     1681 ttcctgcgca agctgagaac gtctgataat ggggcaatcc ctaaccagat ccacgcctat
     1741 gaactggaaa aaatcatcga gaggcaaggg aaggattacc catttctgtt ggaaaataag
     1801 gataagttgc tgagcatatt gacgtttaag atcccctact atgtaggacc cctcgccaag
     1861 gggagtaact cccggttcgc ttggattaag agggcgactt ccagtgacat cctggacgac
     1921 aatgatgagg acacgcggaa tggcaagatt aggccttgga attatcagaa gctgataaat
     1981 atggatgaaa ccagagatgc attcataaca aacctgatcg ggaatgacat tatcctcctg
     2041 aacgaaaagg ttctgccaaa aagatctctt atatacgagg aggtcatgct tcagaacgaa
     2101 ctcactaggg tgaagtataa agacaagtat ggcaaggctc acttcttcga ctccgagctg
     2161 cgacagaata ttattaatgg gctgttcaaa aataacagca agcgagtgaa cgccaagtct
     2221 ctgattaagt acctcagtga caatcataag gacctgaacg caatcgaaat tgtgtccggg
     2281 gtggaaaaag ggaaatcatt caatagtacg ttgaaaacat acaatgacct gaagaccatc
     2341 ttttccgagg aactcttgga tagtgagatt taccagaaag aactggagga gatcatcaaa
     2401 gtcattactg tcttcgacga taaaaagagc attaagaact atctcacaaa attcttcggc
     2461 cacctggaga tcctggacga ggaaaaaatt aatcagctga gcaaactgcg ctattccggc
     2521 tggggaagat acagtgcgaa gctgctcctg gacattcggg atgaagatac cggcttcaat
     2581 ctgctccaat tccttcgaaa cgatgaggag aatcggaacc ttacaaagct gatttccgac
     2641 aatactctga gttttgagcc taaaatcaag gatattcaaa gcaaaagcac tattgaagac
     2701 gatatcttcg acgaaataaa aaagcttgca gggagtccgg ccattaagag gggaatattg
     2761 aactccatca agattgttga tgagctcgtg caaatcatag ggtatcctcc tcataacatc
     2821 gtgatagaga tggcgcggga gaacatgacg accgaagagg gccagaagaa agctaagacc
     2881 agaaaaacta agttggaaag cgcactgaag aacatcgaaa actctctttt ggagaatgga
     2941 aaagtgccac acagcgacga gcagctccag agcgaaaaac tgtatctgta ctacctccag
     3001 aacggaaaag atatgtatac cctcgataaa actggttccc ccgcacccct ctacctcgac
     3061 cagctggatc agtatgaggt tgaccacatc attccataca gttttctgcc aatagattct
     3121 atcgacaaca aagtgcttac ccatcgcgag aataatcagc agaagctgaa caacattcct
     3181 gataaggaaa ccgtggcgaa tatgaaacca ttttgggaaa agctgtacaa cgccaagttg
     3241 atctctcaga caaaatatca gaggctcact acatccgaaa ggaccccaga cggggtactg
     3301 acagagtcca tgaaggccgg attcatcgag aggcagctgg tcgagacgag acagattatt
     3361 aaacacgtgg cccggatcct cgataatcga ttctctgaca ccaaaataat cactctgaag
     3421 agccagctca ttactaactt ccgaaacacc tttcacatcg caaagatccg agagctgaat
     3481 gactaccatc atgctcatga cgcctacttg gccgtggtgg tgggtcagac actcttgaag
     3541 gtgtatccta aactcgcacc agaactcatt tacggccatc acgcgcactt taaccggcac
     3601 gaagagaata aagccacact gagaaagcac ctctactcta atataatgcg gttcttcaat
     3661 aacccagact ccaaagtaag caaggacatt tgggactgta acagagatct gcccatcatc
     3721 aaagatgtga tttataacag tcagataaac ttcgtcaaac ggacaatgat aaaaaagggt
     3781 gctttttaca atcagaatcc agtgggcaag ttcaacaaac aacttgctgc taacaataga
     3841 tatcctctca aaaccaaagc cttgtgcctg gacacatcca tctacggtgg ctatggacca
     3901 atgaacagcg cattgagcat tattatcatc gccgaaagat tcaacgaaaa aaaaggtaag
     3961 atcgaaaccg tcaaagagtt ccatgacatt ttcatcattg attacgagaa attcaacaac
     4021 aatccgtttc agtttctgaa tgacactagc gaaaacggct tcctcaagaa aaataatata
     4081 aatagagtgt tgggattcta tcggatcccc aagtatagtt tgatgcagaa gattgatgga
     4141 actcggatgt tgttcgaatc taaaagcaat ctgcataagg cgactcagtt caaactcact
     4201 aaaacccaga atgagctgtt ctttcacatg aagcgcctgt tgacgaagag taaccttatg
     4261 gacctcaagt ctaagtccgc gattaaagag tcccagaact ttatcctgaa gcataaggaa
     4321 gagtttgaca acatttcaaa ccagctgtcc gcgttcagcc agaagatgct tgggaacacc
     4381 actagcctga aaaacctcat aaaaggatac aatgaacgga agatcaagga aatcgatatc
     4441 cgagacgaaa cgatcaaata tttttacgac aacttcatca aaatgttttc cttcgtcaaa
     4501 agcggcgccc ccaaagatat taacgatttt tttgataata aatgtaccgt agctcgcatg
     4561 aggccaaagc ctgataaaaa gctcctgaac gccacactga ttcaccagtc catcaccggc
     4621 ctgtacgaga ctcggatcga tctctccaag ctgggtgagg atagcagggc tgaccccaag
     4681 aagaagagga aggtgaggtc catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4741 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4801 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4861 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4921 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcgtcttcc
     4981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6901 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6961 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acac
//
