LOCUS       sequence-123991-        1057 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 1057)
  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 1057)
  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..1057
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(262..337)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(433..450)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(433..449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(442..464)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      662..1042
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(749..768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     959..980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atgaccatga ttacgccaag ctccttcctc ttccagccct tcctctttcg ggcggcagca
       61 agatcccttc tgatctaaag tccaaccggc gagagtgaac ggcactgcgc ttggtttagt
      121 tgtcaagaga gaataactct cggacgcgga aggattcgta gcgtatatcg cctattgcgg
      181 aagcgtaacc aagtcttacg accccaaaag aggttgactg aactgggacg cggcactggc
      241 gcaaagcttg gatccaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac
      301 tagccttatt ttaacttgct atttctagct ctaaaaccca tggaattcgt gtcgcatgaa
      361 taacgggcag ttatcaaatg agcacggcga cagcctgtga aagtcctctc cactgactgt
      421 agcctccaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      481 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      541 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      601 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
      661 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      721 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      781 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      841 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
      901 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      961 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     1021 taagggattt tgccgatttc ggtctattgg ttaaaaa
//
