LOCUS       sequence-151822-        1036 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  tRNA and scaffold for the assembly of GBoligomers for the last 
            position (positon [n]) of a polycistronic tRNA-gRNA (2- and 3-part 
            multiplexing).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1036)
  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 1036)
  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 [n] (GB1207)
FEATURES             Location/Qualifiers
     source          1..1036
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    82..106
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(118..193)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      322..331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      326..335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(365..384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(366..384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(391..408)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(391..407)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(400..422)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      548..1003
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(635..654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     845..866
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acgcgttggg agctctccca tatggtcgac ctgcaggcgg ccgcgaattc actagtgatc
       61 ccgtctcgct caagcgtaat gccaactttg tacaagaaag ctgggtctag aaaaaaagca
      121 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctattt
      181 ctagctctaa aactgagacg gcgcgcgccg tctctgcacc agccgggaat cgaacccggg
      241 tctgtaccgt ggcagggtac tattctacca ctagaccact ggtgctttgt tgccgagcga
      301 gacgggaatc gaattcccgc ggccgccatg gcggccggga gcatgcgacg tcgggcccaa
      361 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
      421 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
      481 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
      541 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      601 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      661 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      721 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      781 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      841 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      901 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      961 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     1021 tccttacgca tctgtg
//
