LOCUS       sequence-199458-        2287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a guide RNA (gRNA) to target human IRF-1 promoter for the 
            Sa-Cas9 system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2287)
  AUTHORS   Fujita T, Yuno M, Fujii H
  TITLE     enChIP systems using different CRISPR orthologues and epitope tags.
  JOURNAL   BMC Res Notes. 2018 Feb 27;11(1):154. doi: 
            10.1186/s13104-018-3262-4.
  PUBMED    29482606
REFERENCE   2  (bases 1 to 2287)
  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: "BMC Res 
            Notes."; date: "2018-02-27"; volume: "11(1):154. doi"; pages: " 
            10.1186/s13104-018-3262-4"
COMMENT     Sequence Label: hIRF-1 gRNA #351 (Sa)
FEATURES             Location/Qualifiers
     source          1..2287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(538..1126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(618..637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1297..2088,2089..2157))
                     /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     1920..1939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2158..2262)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc ccagtgggat atcaagaagg gttttagtac
      361 tctgtaatga aaattacaga atctactaaa acaaggcaaa atgccgtgtt tatctcgtca
      421 acttgttggc gagatttttt taagcttggg ccgctcgagg tacctctcta catatgacat
      481 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      541 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      601 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      661 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      721 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      781 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      841 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      901 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
      961 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1021 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1081 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1141 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1201 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1261 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1321 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1381 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1441 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1501 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1561 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1621 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1681 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1741 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1801 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     1861 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1921 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1981 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2041 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2101 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2161 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2221 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2281 gccacct
//
