LOCUS       sequence-100235-         752 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Serves as a shuttle vector for target oligo before insertion into 
            Cas9 destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 752)
  AUTHORS   Michno JM, Wang X, Liu J, Curtin SJ, Kono TJ, Stupar RM
  TITLE     CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a 
            new web-tool and a modified Cas9 enzyme.
  JOURNAL   GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063.
  PUBMED    26479970
REFERENCE   2  (bases 1 to 752)
  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: "GM Crops 
            Food."; date: "2015"; volume: "6(4):243-52. doi"; pages: " 
            10.1080/21645698.2015.1106063"
COMMENT     Sequence Label: pBlu/gRNA
FEATURES             Location/Qualifiers
     source          1..752
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     171..193
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     185..202
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     186..202
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     260..276
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     260..276
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(304..379)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
       61 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      181 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattggagct
      241 ccaccgcggt ggcggccgct ctagaactag tggatccccc gggctgcagg aattcaaaaa
      301 aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg
      361 ctatttctag ctctaaaaca ggtcttctcg aagacccaat cactacttcg actctagctg
      421 tatataaact cagcttcgtt ttcttatcta agcgatgtgg gacttttgaa gattgttttc
      481 aacttaaatg ggcctatata agaaatacta ttgttctttc ccatataaat gggcctgctt
      541 ctcttctttc agattcccag gggccttttg aagattatct tcatatctta agaatgaaga
      601 tgttttattc aatcaaattc ttgaaggttc gatgcctaat cattctaatc ctgggacaaa
      661 ctatgaaaca agatacaaaa actccgaatg gaaagttaaa aagaagaaaa cgaaagctac
      721 ggttcaagaa aatgtaagct gataaacaaa aa
//
