LOCUS       sequence-403566-        2377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of full-length AsCas12f sgRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2377)
  AUTHORS   Wu T, Liu C, Zou S, Lyu R, Yang B, Yan H, Zhao M, Tang W
  TITLE     An engineered hypercompact CRISPR-Cas12f system with boosted 
            gene-editing activity.
  JOURNAL   Nat Chem Biol. 2023 Jul 3. doi: 10.1038/s41589-023-01380-9.
  PUBMED    37400536
REFERENCE   2  (bases 1 to 2377)
  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: "Nat Chem 
            Biol. 2023 Jul 3. doi: 10.1038/s41589-023-01380-9."
COMMENT     Sequence Label: U6-AsCas12f-full-length-sgRNA
FEATURES             Location/Qualifiers
     source          1..2377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     376..395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(614..718)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        825..1073
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     825..845
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     996..1015
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(1371..1959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1451..1470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2130..2377,1..544,545..613))
                     /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"
ORIGIN
        1 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
       61 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      121 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      181 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
      241 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
      301 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
      361 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
      421 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
      481 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
      541 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
      601 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
      661 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
      721 catttccccg aaaagtgcca cctgacgtcg ctagctgtac aaaaaagcag gctttaaagg
      781 aaccaattca gtcgactgga tccggtacca aggtcgggca ggaagagggc ctatttccca
      841 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
      901 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
      961 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     1021 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggattc
     1081 gtcggttcag cgacgataag ccgagaagtg ccaataaaac tgttaagtgg tttggtaacg
     1141 ctcggtaagg tagccaaaag gctgaaactc cgtgcacaaa gaccgcacgg acgcttcaca
     1201 tatagctcat aaacaagggt ttgcgagcta gcttgtggag tgtgaacagg catcactgcc
     1261 ccctgatttt ttttaagctt gggccgctcg aggtacctct ctacatatga catgtgagca
     1321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1801 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2341 ggtcctgcaa ctttatccgc ctccatccag tctatta
//
