LOCUS       sequence-238302-        2034 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses gRNA targeting the IL1RN promoter (SpyCas9 scaffold).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2034)
  AUTHORS   Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A, 
            Waldhauer MC, Borner K, Fakhiri J, Schmelas C, Dietz L, Grimm D, 
            Correia BE, Eils R, Niopek D
  TITLE     Engineered anti-CRISPR proteins for optogenetic control of 
            CRISPR-Cas9.
  JOURNAL   Nat Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi: 
            10.1038/s41592-018-0178-9.
  PUBMED    30377362
REFERENCE   2  (bases 1 to 2034)
  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 
            Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi: 
            10.1038/s41592-018-0178-9."
COMMENT     Sequence Label: IL1RN gRNA1
FEATURES             Location/Qualifiers
     source          1..2034
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        228..476
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     228..248
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     399..418
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             complement(join(661..1374,1375..1377,1378..1380))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(664..1374,1375..1377,1378..1380))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(706..727)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1055..1074
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1314..1335
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1377..1386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1424..1685)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     1962..1981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgagtccaac acccgtggga atcccatggg caccatggcc cctcgctcca aaaatgcttt
       61 cgcgtcgcgc agacactgct cggtagtttc ggggatcagc gtttgagtaa gagcccgcgt
      121 ctgaaccctc cgcgccgccc cggccccagt ggaaagacgc gcaggcaaaa cgcaccacgt
      181 gacggagcgt gaccgcgcgc cgagcgcgcg ccaaggtcgg gcaggaagag ggcctatttc
      241 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attagaatta
      301 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
      361 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
      421 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacacctgta
      481 ctctctgagg tgctcgttta agagctatgc tggaaacagc atagcaagtt taaataaggc
      541 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgc ggccgcctcg
      601 agtgatcaaa aaaccaacac acgcttccaa tgaaaataaa cgatccttta ttgctagcct
      661 ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
      721 gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta
      781 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
      841 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc
      901 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
      961 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1021 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1081 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1141 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1201 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1261 gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1321 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1381 ggtggcgacc ggtggatccc gggcccgcgg tacccagctt ggaggtgcac accaatgtgg
     1441 tgaatggtca aatggcgttt attgtatcga gctaggcact taaatacaat atctctgcaa
     1501 tgcggaattc agtggttcgt ccaatccatg tcagacccgt ctgttgcctt cctaataagg
     1561 cacgatcgta ccaccttact tccaccaatc ggcatgcacg gtgctttttc tctccttgta
     1621 aggcatgttg ctaactcatc gttaccatgt tgcaagacta caagagtatt gcataagact
     1681 acatttcccc ctccctatgc aaaagcgaaa ctactatatc ctgaggggac tcctaaccgc
     1741 gtacaaccga agccccgctt ttcgcctaaa cacaccctag tcccctcaga tacgcgtata
     1801 tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca
     1861 attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac
     1921 ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt
     1981 actgagagtg caccatacgg atctgcgatg ataagctgtc aaacatgaga attg
//
