LOCUS       sequence-307476-        2045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  End-linker for cloning 4 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: pDGE510
FEATURES             Location/Qualifiers
     source          1..2045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     183..200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(330..348)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        416..520
                     /gene="bla"
                     /label=AmpR promoter
     CDS             521..1315
                     /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(1486..2045,1..29)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1975..1994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
       61 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      181 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
      241 cgcgttggcc gattcattaa tcactctgtg gtctcaccag ataagcttta gctttgagac
      301 cacgaagtgg ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      361 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      421 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      481 accctgataa atgcttcaat aatattgaaa aaggaagagt atggctaaaa tgagaatatc
      541 accggaattg aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc
      601 tcctgctaag gtatataagc tggtgggaga aaatgaaaac ctatatttaa aaatgacgga
      661 cagccggtat aaagggacca cctatgatgt ggaacgggaa aaggacatga tgctatggct
      721 ggaaggaaag ctgcctgttc caaaggtcct gcactttgaa cggcatgatg gctggagcaa
      781 tctgctcatg agtgaggccg atggcgtcct ttgctcggaa gagtatgaag atgaacaaag
      841 ccctgaaaag attatcgagc tgtatgcgga gtgcatcagg ctctttcact ccatcgacat
      901 atcggattgt ccctatacga atagcttaga cagccgctta gccgaattgg attacttact
      961 gaataacgat ctggccgatg tggattgcga aaactgggaa gaggacactc catttaaaga
     1021 tccgcgcgag ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc
     1081 ccacggcgac ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat
     1141 tgatcttggg agaagcggca gggcggacaa gtggtatgac attgccttct gcgtccggtc
     1201 gctcagggag gatatcgggg aagaacagta tgtcgagcta ttttttgact tactggggat
     1261 caagcctgat tgggagaaaa taaaatatta tattttactg gatgaattgt tttagctgtc
     1321 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1381 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1441 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1501 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1561 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1621 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1681 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1741 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1801 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1861 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1921 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1981 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2041 gtgat
//
