LOCUS       sequence-403570-        2305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of trancated AsCas12f sgRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2305)
  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 2305)
  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-sgRNA-v2
FEATURES             Location/Qualifiers
     source          1..2305
                     /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(1299..1887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1379..1398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2058..2305,1..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 tagccttcgg gcaagaccac tgaacaggca tcactgcccc ctgatttttt
     1201 ttaagcttgg gccgctcgag gtacctctct acatatgaca tgtgagcaaa aggccagcaa
     1261 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1321 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1381 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1441 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1501 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1561 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1621 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1681 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1741 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1801 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1861 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1921 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1981 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2041 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2101 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2161 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2221 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2281 ttatccgcct ccatccagtc tatta
//
