LOCUS       sequence-151825-        1046 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  tRNA and scaffold for the assembly of GBoligomers for the first 
            position (positon [D1_n-1]) of a polycistronic tRNA-gRNA regulated 
            by the U6-26 or U6-1 promoter (2-part multiplexing).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1046)
  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 1046)
  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_n-1] (GB1208)
FEATURES             Location/Qualifiers
     source          1..1046
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      276..285
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      280..289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(319..338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(320..338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(345..362)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(345..361)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(354..376)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      502..957
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(589..608)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     799..820
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgttgggagc tctcccatat ggtcgacctg caggcggccg cgaattcact agtgatcccg
       61 tctcgctcga ttgaacaaag caccagtggt ctagtggtag aatagtaccc tgccacggta
      121 cagacccggg ttcgattccc ggctggtgca gagacggcgc gcgccgtctc agttttagag
      181 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
      241 tcggcaatga gcgagacggg aatcgaattc ccgcggccgc catggcggcc gggagcatgc
      301 gacgtcgggc ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
      361 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      421 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      481 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      781 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      841 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      901 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
      961 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatca ggtggcactt
     1021 ttcggggaaa tgtgcgcgga acccct
//
