LOCUS       sequence-378974-        4460 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  vector for encoding a human codon-optimized Cas12Max driven by CBh 
            promoter, guide RNAs compatible with xCas12i driven by hU6, and 
            mCherry driven by CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4460)
  AUTHORS   Zhang H, Kong X, Xue M, Hu J, Wang Z, Wei Y, Wang H, Zhou J, Zhang 
            W, Xu M, Shen X, Yin F, Ai Z, Huang G, Xia J, Song X, Li H, Yuan Y, 
            Li J, Zhong N, Zhang M, Zhou Y, Yang H
  TITLE     An engineered xCas12i with high activity, high specificity, and 
            broad PAM range.
  JOURNAL   Protein Cell. 2023 Jun 28;14(7):538-543. doi: 
            10.1093/procel/pwac052.
  PUBMED    37378659
REFERENCE   2  (bases 1 to 4460)
  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"; doi: "10"; journalName: 
            "Protein Cell"; date: "2023-06-28- 28"; volume: "14"; issue: "7"; 
            pages: "538-543"
COMMENT     Sequence Label: Cas12Max
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        1..286
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        288..565
                     /label=chicken beta-actin promoter
     intron          566..794
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      806..815
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             815..880
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             887..907
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4172..4219
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4247..4264)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4253..4460
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      121 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      181 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
      241 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      301 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      361 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg
      421 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      481 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      541 taaaaagcga agcgcgcggc gggcgggagt cgctgcgacg ctgccttcgc cccgtgcccc
      601 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
      661 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gctgagcaag aggtaagggt
      721 ttaagggatg gttggttggt ggggtattaa tgtttaatta cctggagcac ctgcctgaaa
      781 tcactttttt tcaggttgga ccggtgccac catggactat aaggaccacg acggagacta
      841 caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg
      901 gaaggtcggt atccacggag tcccagcagc catgagttct gatgtggtgc ggccttataa
      961 cacaaagctg ctcccagata acagaaagca caatatgttc ctgcagacct tcaagcggct
     1021 gaacagcatc tctctgaacc acttcgacct gctgatctgc ctgtacgctg caatcaccaa
     1081 caagaaggcc gaggaataca agtctgaaaa ggaagcccac gtgaccgccg atagcctgtg
     1141 tgccatcaat tggttcagac ccatgagcaa gagatacagc aaatacgcca ccaccacctt
     1201 caacatgtta gaactgttta aggagtacag cggccacgag cctgatgcct attccaagaa
     1261 ctacctgatg agcaatatcg acagcgacag attcgtgtgg gtggattgta ggaagttcgc
     1321 taaggacttt gcctatcaga tggaactggg tttccacgag ttcaccgtgt tggccgaaac
     1381 cctgctggct aattctatcc tggtgctgaa cgagagcacc aaggccaatt gggcttgggg
     1441 aaccgtgtct gccctgtacg gcggcggaga taaggaggac agcacactga agagcaagat
     1501 tctgctggcc ttcgtggacg ccctgaacaa ccacgagctg aaaacaaaga gagaaatctt
     1561 gaatcaagtg tgtgaatctc tgaaatacca gagctaccag gacatgtacg tggattttag
     1621 aagcgtggtt gacgaaaacg gcaacaagaa gtctcctaga ggctctatgc ctatcgtgac
     1681 caagttcgag acagacgacc tgatcagcga caaccaaaga aaggccatga tcagcaactt
     1741 cactaagaac gccgctgcca aggcagctaa gaaacctatc ccttacttgg accgcctgaa
     1801 ggagcacatg gtgtccctgt gcgacgagta caatgtgtat gcctgggccg cggccatcac
     1861 aaacagcaac gccgacgtga ccgcccggaa taccagaaac ctgacattca tcggcgaaca
     1921 gaacagcaga cgaaaggaac tgagcgtgct gcagacaaca accaacgaga aggctaagga
     1981 catcctgaac aagatcaacg acaacctgat tcaggaggtg cggtacaccc ctgcccctaa
     2041 gcacctgggc agagatctgg ccaacctgtt tgatacactg aaggaaaagg acatcaacaa
     2101 catcgagaac gaagaagaga aacagaacgt gatcaatgac tgtatcgagc agtacgtgga
     2161 cgattgcaga agcctcaacc ggaaccccat cgcagccctc ctgaagcaca tctctaggta
     2221 ctacgaggat ttcagcgcca agaatttcct ggacggcgcc aagctgaacg tgctgactga
     2281 ggtggtgaac cggcagaagg cccaccccac catctggagc gagaaggctt acacctggat
     2341 cagcaagttc gacaagaacc ggagacaggc caacagcagc ctggtcggat gggttgtgcc
     2401 ccccgaggag gtgcacaagg agaaaatcgc cggacagcag agcatgatgt gggtgaccct
     2461 caccctgctg gacgacggca agtgggtcaa acatcacatc cccttcagcg acagcagata
     2521 ctacagcgaa gtgtacgcct acaaccctaa tctgccttat ctggacggag gcatcccaag
     2581 acagagcaag ttcggcaaca aaccaacaac caacctgaca gccgagtccc aggccctcct
     2641 ggctaattct aagtacaaga aagccaacaa gagcttcctg cgggctaaag agaatgccac
     2701 acacaacgtg cgggtgtccc ctaacacctc tctgtgcatt agactgctga aggacagcgc
     2761 cggaaaccag atgttcgaca aaatcggcaa cgtgctcttc ggcatgcaga tcaaccacaa
     2821 gatcaccgtg ggaaaaccta actacaagat cgaggtgggc gacagattcc tgggcttcga
     2881 tcagaaccag agcgagaacc acacctacgc cgtgctgcag agagtgtccg agagcagtca
     2941 cgacacccac cactttaacg gctgggacgt gaaggtgctg gaaaagggca aagtgaccag
     3001 cgatgtgatc gtgcgggacg aggtctacga ccaactgtct tacgagggcg tcccctacga
     3061 tagcagcaag ttcgccgagt ggcgggacaa gcgcagaaga tttgtgcttg agaacctgag
     3121 catccagctg gaagagggca agaccttcct gacagagttc gacaagctga ataaggacag
     3181 cctgtaccgc tggaacatga actacctgaa actgctgaga aaggccatcc gggccggagg
     3241 caaagagttc gccaagatcg ctaagacaga gatcttcgag ctggcggtgg aaagattcgg
     3301 ccctattaac ctgggcagcc tgtcccagat cagccttaag atgattgcct cctttaaggg
     3361 cgtggtccag tcctacttct ccgtgagcgg ctgcgtggat gatgcctcca aaaaggccca
     3421 tgattctatg ctgttcacat ttatgtgcgc cgccgaagaa aagcggacca acaagagaga
     3481 agaaaagacc aacagagccg ccagctttat cctgcaaaaa gcctacctgc atggctgcaa
     3541 gatgatcgtg tgcgaggacg accttcctgt ggccgacggc aagacaggca aagcccagaa
     3601 tgccgaccgg atggactggt gcgccagagc cctggccaag aaggtgaacg acggctgtgt
     3661 tgccatgagc atctgctaca gagctatccc tgcctacatg agcagccacc aggacccctt
     3721 tgtgcacatg caggataaga aaaccagcgt gctgcggcct agattcatgg aagttaataa
     3781 ggatagcatc agagactacc acgtggcggg cctgagaaga atgctgaaca gcaagagtga
     3841 cgctggcacc agtgtttatt accggcaagc tgccctgcat ttctgcgaag ccctgggcgt
     3901 gagccctgaa ctggtgaaaa acaagaaaac ccacgccgcc gaactgggca agcacatggg
     3961 cagcgctatg ctgatgccct ggagaggcgg tagagtgtac atcgccagca aaaagctgac
     4021 ctccgatgcc aaatcagtga agtactgcgg cgaggatatg tggcagtacc acgccgatga
     4081 gatcgccgct gttaacatcg ccatgtatga ggtgtgctgc cagaccggcg ctttcggaaa
     4141 gaaacagaaa aaatcggacg agctgcctgg aaaaaggccg gcggccacga agaaggccgg
     4201 ccaggcaaag aagaagaagt aagaattcct agagctcgct gatcagcctc gactgtgcct
     4261 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4321 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4381 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     4441 aatagcaggc atgctgggga
//
