LOCUS       sequence-151793-         909 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  tRNA and scaffold for the assembly of GBoligomers for the first 
            position (positon [D1_2]) of a polycistronic tRNA-gRNA regulated by 
            the dicot U6-26 or U6-1 promoter (3-part multiplexing).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 909)
  AUTHORS   Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo 
            P, Blanca J, Granell A, Orzaez D
  TITLE     A modular toolbox for gRNA-Cas9 genome engineering in plants based 
            on the GoldenBraid standard.
  JOURNAL   Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. 
            eCollection 2016.
  PUBMED    26839579
REFERENCE   2  (bases 1 to 909)
  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: "Plant 
            Methods."; date: "2016-02-1"; volume: "12:10. doi"; pages: " 
            10.1186/s13007-016-0101-2. eCollection 2016"
COMMENT     Sequence Label: tRNA-gRNA position [D1_2] (GB1205)
FEATURES             Location/Qualifiers
     source          1..909
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(23..98)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(293..311)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(294..311)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(329..345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(329..345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(342..364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    353..369
                     /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(377..383,384..401,402..407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    422..443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(560..577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(811..830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattcgattc ccgtctcgct cagcaccgac tcggtgccac tttttcaagt tgataacgga
       61 ctagccttat tttaacttgc tatttctagc tctaaaactg agacggcgcg cgccgtctct
      121 gcaccagccg ggaatcgaac ccgggtctgt accgtggcag ggtactattc taccactaga
      181 ccactggtgc tttgttcaat cgagcgagac gggatcacta gtgaattcgc ggccgcctgc
      241 aggtcgacca tatgggagag ctcccaacgc gttggatgca tagcttgagt attctatagt
      301 gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      361 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      421 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      481 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      541 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      901 cttcgggaa
//
