LOCUS       sequence-152161-        1015 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  tRNA and scaffold for the assembly of GBoligomers for the first 
            position (positon [M1_n-1]) of a polycistronic tRNA-gRNA regulated 
            by the (monocot) U3 promoter (2-part multiplexing).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1015)
  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 1015)
  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 [M1_n-1] (GB1210)
FEATURES             Location/Qualifiers
     source          1..1015
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      277..286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      281..290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(320..339)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(321..339)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(346..363)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(346..362)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(355..377)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      503..958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(590..609)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     800..821
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgttgggag ctctcccata tggtcgacct gcaggcggcc gcgaattcac tagtgatccc
       61 gtctcgctca ttgccgactc ggtgccactt tttcaagttg ataacggact agccttattt
      121 taacttgcta tttctagctc taaaactgag acggcgcgcg ccgtctctgc accagccggg
      181 aatcgaaccc gggtctgtac cgtggcaggg tactattcta ccactagacc actggtgctt
      241 tgtttgcccg agcgagacgg gaatcgaatt cccgcggccg ccatggcggc cgggagcatg
      301 cgacgtcggg cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
      361 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
      421 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
      481 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      541 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      601 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      661 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      721 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      781 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      841 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      901 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
      961 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatc aggtg
//
