LOCUS       sequence-490427-        2274 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 2274)
  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 2274)
  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-mini-sgRNA
FEATURES             Location/Qualifiers
     source          1..2274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..270
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     22..42
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     193..212
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(465..1053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(545..564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1224..2015,2016..2084))
                     /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     1847..1866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2085..2189)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
       61 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      121 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      181 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      241 ttatatatct tgtggaaagg acgaaacacc ggtgggaggt ctgtccccac catggggtcc
      301 aaacgactaa ccatgcattg gaaagtgcat ggagcagaag aaacacagta ggagggactt
      361 ttttttttaa gcttgggccg ctcgaggtac ctctctacat atgacatgtg agcaaaaggc
      421 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      481 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      541 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      601 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      661 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      721 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      781 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      841 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      901 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      961 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1021 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1081 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1141 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1201 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1261 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1321 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1381 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1441 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1501 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1561 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1621 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1681 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1741 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1801 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1861 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1921 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1981 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2041 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2101 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2161 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     2221 gctagctgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg atcc
//
