LOCUS       sequence-140157-        1957 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid constitutively expresses gRNA-r10 from an AHD1 promoter
            using the RGR design. Integrates in the W303 HIS locus and restores 
            HIS function..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1957)
  AUTHORS   Gander MW, Vrana JD, Voje Jr WE, Carothers JM, Klavins E
  TITLE     Robust digital logic circuits in eukaryotic cells with CRISPR/dCas9 
            NOR gates
  JOURNAL   bioRxiv 041871
REFERENCE   2  (bases 1 to 1957)
  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: "bioRxiv 
            041871"
COMMENT     Sequence Label: pMOD8A-RGR-r10
FEATURES             Location/Qualifiers
     source          1..1957
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        741..1445
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     primer_bind     1462..1478
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1462..1478
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     ncRNA           1491..1527
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        1548..1623
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           1628..1695
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     complement(1697..1713)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1727..1745)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     terminator      1757..1946
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
ORIGIN
        1 aagaaatgat ggtaaatgaa ataggaaatc aaggagcatg aaggcaaaag acaaatataa
       61 gggtcgaacg aaaaataaag tgaaaagtgt tgatatgatg tatttggctt tgcggcgccg
      121 aaaaaacgag tttacgcaat tgcacaatca tgctgactct gtggcggacc cgcgctcttg
      181 ccggcccggc gataacgctg ggcgtgaggc tgtgcccggc ggagtttttt gcgcctgcat
      241 tttccaaggt ttaccctgcg ctaaggggcg agattggaga agcaataaga atgccggttg
      301 gggttgcgat gatgacgacc acgacaactg gtgtcattat ttaagttgcc gaaagaacct
      361 gagtgcattt gcaacatgag tatactagaa gaatgagcca agacttgcga gacgcgagtt
      421 tgccggtggt gcgaacaata gagcgaccat gaccttgaag gtgagacgcg cataaccgct
      481 agagtacttt gaagaggaaa cagcaatagg gttgctacca gtataaatag acaggtacat
      541 acaacactgg aaatggttgt ctgtttgagt acgctttcaa ttcatttggg tgtgcacttt
      601 attatgttac aatatggaag ggaactttac acttctccta tgcacatata ttaattaaag
      661 tccaatgcta gtagagaagg ggggtaacac ccctccgcgc tcttttccga tttttttcta
      721 aaccgtggaa tatttcggat atccttttgt tgtttccggg tgtacaatat ggacttcctc
      781 ttttctggca accaaaccca tacatcggga ttcctataat accttcgttg gtctccctaa
      841 catgtaggtg gcggagggga gatatacaat agaacagata ccagacaaga cataatgggc
      901 taaacaagac tacaccaatt acactgcctc attgatggtg gtacataacg aactaatact
      961 gtagccctag acttgatagc catcatcata tcgaagtttc actacccttt ttccatttgc
     1021 catctattga agtaataata ggcgcatgca acttcttttc tttttttttc ttttctctct
     1081 cccccgttgt tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga cactaaagga
     1141 aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta
     1201 tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca
     1261 acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt
     1321 gctatcaagt ataaatagac ctgcaattat taatcttttg tttcctcgtc attgttctcg
     1381 ttccctttct tccttgtttc tttttctgca caatatttca agctatacca agcatacaat
     1441 caactatctc atatacagga ttctagaact agtggatcta caaatgtcac ctgatgagtc
     1501 cgtgaggacg aaacgagtaa gctcgtcgtg acataaacat tcgactcgtt ttagagctag
     1561 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     1621 tgcttttggc cggcatggtc ccagcctcct cgctggcgcc ggctgggcaa catgcttcgg
     1681 catggcgaat gggactgata ccgtcgacct cgagtcaatt agttatgtca cgcttacatt
     1741 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     1801 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     1861 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     1921 ttgagaaggt tttgggacgc tcgaaggctt taatttg
//
