LOCUS       sequence-296162-        2866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For sgRNA expression in monocotyledons protoplasts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2866)
  AUTHORS   Lin Q, Zong Y, Xue C, Wang S, Jin S, Zhu Z, Wang Y, Anzalone AV, 
            Raguram A, Doman JL, Liu DR, Gao C
  TITLE     Prime genome editing in rice and wheat.
  JOURNAL   Nat Biotechnol. 2020 May;38(5):582-585. doi: 
            10.1038/s41587-020-0455-x. Epub 2020 Mar 16.
  PUBMED    32393904
REFERENCE   2  (bases 1 to 2866)
  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"; doi: 
            "10.1038/s41587-020-0455-x"; journalName: "Nat Biotechnol"; date: 
            "2020-05"; volume: "38"; issue: "5"; pages: "582-585"
COMMENT     Sequence Label: QPM-sgR (pTaU3)
FEATURES             Location/Qualifiers
     source          1..2866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        425..500
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     511..527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     511..527
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    540..714
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        859..963
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2031..2619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2520..2539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2773..2790
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggcggcaggg agagttttaa cattgactag cgtgctgata atttgtgaga aataataatt
       61 gacaagtaga tactgacatt tgagaagagc ttctgaactg ttattagtaa caaaaatgga
      121 aagctgatgc acggaaaaag gaaagaaaaa gccatacttt tttttaggta ggaaaagaaa
      181 aagccatacg agactgatgt ctctcagatg ggccgggatc tgtctatcta gcaggcagca
      241 gccctaccaa cctcacgggc cagcaattac gagtccttct aaaacgtccc gccgagggcg
      301 cgtggccgtg ctgtgcagca gcacgtctaa cattagtccc acctcgccag tttacaggga
      361 gcagaaccag cttataagcg gaggcgcggc accaagaagc gagagacgga gtcgattcgt
      421 ctcggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      481 aagtggcacc gagtcggtgc tttttttttt caggaaacag ctatgaccca tggctcgagt
      541 ttctccataa taatgtgtga gtagttccca gataagggaa ttagggttcc tatagggttt
      601 cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc
      661 tatcaataaa atttctaatt cctaaaacca aaatccagta ctaaaatcca gatcccccga
      721 attagagctc gcgcgtagtc ctcggtaata tcgcagaaca aaagtacctg atatcgagtg
      781 tacttcaagt cacaccggcg agtgactagc aggtgatgaa aagccttagg gtcaggtggc
      841 acttttcggg gaaatgtgcg atgaacccct atttgtttat ttttctaaat acattcaaat
      901 atgtatccgc taatgagaca ataaccctga taaatgcttc aattatattg aaaaaggaag
      961 agtatgagta tccaacattt ccgtgtcgcc cttattccct tttttgcggc cttctgcctt
     1021 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1081 gcccgagtgg gttacattga actggatctc aacagcggca aaatcttaga gagttttcgc
     1141 cccgaagaac gcttcccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtgtta
     1201 tcccgtattg atgctgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1261 ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1321 ttatgtagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1381 attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1441 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1501 atgcctgtag caatggcaac aactctccgc aaactattaa ctggcgaact acttactctt
     1561 gcttcacgcc agcaactcat tgactggatg gaggcggaca aagttgcagg accacttctg
     1621 cgctcggcac ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggc
     1681 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1741 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1801 gcctcactga ttaagcattg gtaactgtca gacgaaggga aataaatagt agcccgcctg
     1861 atgtgcgggc ttttttttgg tggtggttac cagtgtcgct tagctaccgt aggtcctctt
     1921 aggcggtccg attagcgaca tagtcatcaa tgccttgacg gcaaaatccc ttaacgtgag
     1981 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2041 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2101 tgtttgccgg atcaagagct actaactctt tttccgaagg taactggctt cagcagagcg
     2161 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2221 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2281 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2341 tcgggctgaa cggggggttc gtgctcacag cccagcttgg agcgaacgac ctacaccgaa
     2401 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2461 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2521 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2581 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2641 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2701 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2761 acgaccgagc gcagcgagtc agtgagcgag gaagcgggag agcgcccacc tgccagggca
     2821 gcgaagagca agccagataa cagtatgcgt atttgcgcgc ttcgaa
//
