LOCUS       sequence-296712-        3499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate sgRNA array cloning vector position 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3499)
  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 3499)
  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: pDGE506
FEATURES             Location/Qualifiers
     source          1..3499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     199..216
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        310..412
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             413..1072
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(479..498)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1414..1719
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1598..1617
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(1800..1818)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1886..1990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1991..2785
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      join(2956..3499,1..45)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3445..3464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
       61 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      121 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      181 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      241 aaaccgcctc tccccgcgcg ttggccgatt cattaatcac tctgtgaaga catgactttg
      301 ctgagacctt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
      361 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
      421 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      481 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      541 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      601 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt
      661 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      721 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
      781 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      841 tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
      901 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
      961 gctgatgccg ctggcgattc aggttcatca tgccgtctgt gatggcttcc atgtcggcag
     1021 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aatcgcgtgg
     1081 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat
     1141 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtgtgc tatgaagcag
     1201 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     1261 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     1321 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     1381 gctcttttgc tgacgagaac agggactggt gaaatgcagt ttaaggttta cacctataaa
     1441 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcctggg
     1501 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     1561 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     1621 agtgtgccgg tttccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     1681 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac
     1741 agccagtcgg tctcagcttc cagttgtctt cacgaagtgg ctcttcagtg gacgaaaggg
     1801 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1861 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1921 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1981 aaggaagagt atggctaaaa tgagaatatc accggaattg aaaaaactga tcgaaaaata
     2041 ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc tggtgggaga
     2101 aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca cctatgatgt
     2161 ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc caaaggtcct
     2221 gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg atggcgtcct
     2281 ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc tgtatgcgga
     2341 gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga atagcttaga
     2401 cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg tggattgcga
     2461 aaactgggaa gaggacactc catttaaaga tccgcgcgag ctgtatgatt ttttaaagac
     2521 ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca gcaacatctt
     2581 tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca gggcggacaa
     2641 gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg aagaacagta
     2701 tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa taaaatatta
     2761 tattttactg gatgaattgt tttagctgtc agaccaagtt tactcatata tactttagat
     2821 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2881 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3061 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3481 tcgccacctc tgacttgag
//
