LOCUS       sequence-307474-        2045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  End-linker for cloning 3 sgRNA arrays into any recipient vector .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2045)
  AUTHORS   Stuttmann J, Barthel K, Martin P, Ordon J, Erickson JL, Herr R, 
            Ferik F, Kretschmer C, Berner T, Keilwagen J, Marillonnet S, Bonas U
  TITLE     Highly efficient multiplex editing: One-shot generation of 8x 
            Nicotiana benthamiana and 12x Arabidopsis mutants.
  JOURNAL   Plant J. 2021 Feb 12. doi: 10.1111/tpj.15197.
  PUBMED    33577114
REFERENCE   2  (bases 1 to 2045)
  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 J. 
            2021 Feb 12. doi: 10.1111/tpj.15197."
COMMENT     Sequence Label: pDGE509
FEATURES             Location/Qualifiers
     source          1..2045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(326..344)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        412..516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             517..1311
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      join(1482..2045,1..25)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1971..1990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
       61 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      121 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      181 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
      241 ttggccgatt cattaatcac tctgtggtct cagactataa gctttagctt tgagaccacg
      301 aagtggctct tcagtggacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      361 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      421 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      481 tgataaatgc ttcaataata ttgaaaaagg aagagtatgg ctaaaatgag aatatcaccg
      541 gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct
      601 gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc
      661 cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa
      721 ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg
      781 ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct
      841 gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg
      901 gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat
      961 aacgatctgg ccgatgtgga ttgcgaaaac tgggaagagg acactccatt taaagatccg
     1021 cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac
     1081 ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat
     1141 cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgctc
     1201 agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag
     1261 cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta gctgtcagac
     1321 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1381 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1621 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2041 tgctc
//
