LOCUS       sequence-322073-        2243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Version of the native Cas9-sgRNA with one native WT aptamer sequence
            and F6 aptamer sequence recognized for Ms2 coat protein, in 3'..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2243)
  AUTHORS   Selma S, Bernabe-Orts JM, Vazquez-Vilar M, Diego-Martin B, Ajenjo M,
            Garcia-Carpintero V, Granell A, Orzaez D
  TITLE     Strong gene activation in plants with genome-wide specificity using 
            a new orthogonal CRISPR/Cas9-based programmable transcriptional 
            activator.
  JOURNAL   Plant Biotechnol J. 2019 Sep;17(9):1703-1705. doi: 
            10.1111/pbi.13138. Epub 2019 May 23.
  PUBMED    31034138
REFERENCE   2  (bases 1 to 2243)
  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: "10.1111/pbi.13138";
            journalName: "Plant Biotechnol J"; date: "2019-09"; volume: "17"; 
            issue: "9"; pages: "1703-1705"
COMMENT     Sequence Label: pUPD2 sgRNA sCRNA 2.0 (GB2461)
FEATURES             Location/Qualifiers
     source          1..2243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        21..96
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        103..121
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      200..257
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(281..298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(452..1040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(532..551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1223..1317)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1338..1997)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1912..1931
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2159..2177
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2181..2224)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(2227..2243,1..5)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tagagcgatg agtggtctcg gttttagagc tagaaatagc aagttaaaat aaggctagtc
       61 cgttatcaac ttgaaaaagt ggcaccgagt cggtgcggga gcacatgagg atcacccatg
      121 tgcgactccc acagtcactg gggagtcttc ccttttttcg cttgagacca ctcatcgcga
      181 ctagtagcgg ccgctgcagt ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt
      241 ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc
      301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
      361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
      421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc cctgacgagc
      481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1141 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1201 ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     1261 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     1321 gcgagctcga tatcaaatta cgccccgccc tgccactcgt cgcagtactg ttgtaattca
     1381 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     1441 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     1501 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     1561 gatacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     1621 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     1681 cagagtgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     1741 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     1801 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     1861 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     1921 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     1981 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     2041 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc
     2101 aactcgagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     2161 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
     2221 tttctggaat tcgcggccgc ttc
//
