LOCUS       sequence-307438-        2045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  End-linker for cloning 2 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: pDGE508
FEATURES             Location/Qualifiers
     source          1..2045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     252..269
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(399..417)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        485..589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             590..1384
                     /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(1555..2045,1..98)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(2044..2045,1..18)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
       61 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      121 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
      181 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      241 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      301 ctctccccgc gcgttggccg attcattaat cactctgtgg tctcacggta taagctttag
      361 ctttgagacc acgaagtggc tcttcagtgg acgaaagggc ctcgtgatac gcctattttt
      421 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      481 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      541 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tggctaaaat
      601 gagaatatca ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga
      661 aggaatgtct cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa
      721 aatgacggac agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat
      781 gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg
      841 ctggagcaat ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga
      901 tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc
      961 catcgacata tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga
     1021 ttacttactg aataacgatc tggccgatgt ggattgcgaa aactgggaag aggacactcc
     1081 atttaaagat ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact
     1141 tgtcttttcc cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag
     1201 tggctttatt gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg
     1261 cgtccggtcg ctcagggagg atatcgggga agaacagtat gtcgagctat tttttgactt
     1321 actggggatc aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt
     1381 ttagctgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1441 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1501 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1561 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1621 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1681 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1741 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1801 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1861 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1921 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1981 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2041 agggg
//
