LOCUS       sequence-490432-        2379 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  for HEK293T human cell genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2379)
  AUTHORS   Jin S, Zhu Z, Li Y, Zhang S, Liu Y, Li D, Li Y, Luo Y, Cheng Z, Zhao
            KT, Gao Q, Yang G, Li H, Liang R, Zhang R, Qiu JL, Zhang YE, Liu JG,
            Gao C
  TITLE     Functional RNA splitting drove the evolutionary emergence of type V 
            CRISPR-Cas systems from transposons.
  JOURNAL   Cell. 2025 Sep 29:S0092-8674(25)01035-9. doi: 
            10.1016/j.cell.2025.09.004.
  PUBMED    41027434
REFERENCE   2  (bases 1 to 2379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2025 
            Sep 29:S0092-8674(25)01035-9. doi: 10.1016/j.cell.2025.09.004."
COMMENT     Sequence Label: phU6-LaTranC-sgRNA
FEATURES             Location/Qualifiers
     source          1..2379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        76..324
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     76..96
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     247..266
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(624..1212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(704..723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1383..2174,2175..2243))
                     /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     2006..2025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2244..2348)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gctagctgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg atccggtacc
       61 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
      121 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
      181 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      241 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      301 atcttgtgga aaggacgaaa caccggtggg aggtctgtcc ccaccatggg gtgcgaacct
      361 tgtgtgctca tcattgccgt gagcgttcgc acgtccaaac gaccatatca tcgttgccct
      421 gcgaccattc agcggcaatc aagacgcagg catgatatgt aaccatgcat tggaaagtgc
      481 atggagcaga agaaacacag taggagggac ttacgttagt tttttttaag cttgggccgc
      541 tcgaggtacc tctctacata tgacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      601 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      661 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      721 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      781 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      841 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      901 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      961 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1021 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1081 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1141 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1201 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1261 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1321 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1381 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1441 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1501 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1561 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1621 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1681 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1741 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1801 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1861 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1921 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1981 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2041 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2101 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2161 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2221 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2281 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2341 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
