LOCUS       sequence-123990-        1095 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contains the sgRNA backbone sequence (tracrRNA, 82 bp) and is used 
            as DNA template to amplify a specific sgRNA using a forward primer 
            with the protospacer sequence for gene targeting..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1095)
  AUTHORS   Lander N, Li ZH, Niyogi S, Docampo R
  TITLE     CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 
            Genes in Trypanosoma cruzi Reveals Their Role in Flagellar 
            Attachment.
  JOURNAL   MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15.
  PUBMED    26199333
REFERENCE   2  (bases 1 to 1095)
  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: "MBio."; 
            date: "2015-07-21"; volume: "6(4). pii: e01012-15. doi"; pages: " 
            10.1128/mBio.01012-15"
COMMENT     Sequence Label: pUC_sgRNA
FEATURES             Location/Qualifiers
     source          1..1095
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        96..171
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(428..444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(428..444)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(441..463)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    452..468
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(476..482,483..500,501..506))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    521..542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(659..676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(910..929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaattggagg ctacagtcag tggagaggac tttcacaggc tgtcgccgtg ctcatttgat
       61 aactgcccgt tattcatgcg acacgaattc catgggtttt agagctagaa atagcaagtt
      121 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgga
      181 tccaagcttt gcgccagtgc cgcgtcccag ttcagtcaac ctcttttggg gtcgtaagac
      241 ttggttacgc ttccgcaata ggcgatatac gctacgaatc cttccgcgtc cgagagttat
      301 tctctcttga caactaaacc aagcgcagtg ccgttcactc tcgccggttg gactttagat
      361 cagaagggat cttgctgccg cccgaaagag gaagggctgg aagaggaagg agcttggcgt
      421 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      481 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      541 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1081 gtgtgcacga acccc
//
