LOCUS       sequence-307433-        3502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate sgRNA array cloning vector position 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3502)
  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 3502)
  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: pDGE505
FEATURES             Location/Qualifiers
     source          1..3502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..25
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     259..276
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        371..473
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             474..1157
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(540..559)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1477..1782
                     /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     1661..1680
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(1863..1881)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1949..2053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2054..2848
                     /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(3019..3502,1..105)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
       61 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      121 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      181 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      241 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      301 aaaccgcctc tccccgcgcg ttggccgatt cattaatcac tcctgtgaag acatcggttt
      361 gctgagacct tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
      421 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
      481 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
      541 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
      601 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
      661 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
      721 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
      781 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
      841 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
      901 ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
      961 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     1021 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1081 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggggggcgt aaacgccgcg
     1141 tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg
     1201 tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag
     1261 cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg
     1321 tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg
     1381 aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga
     1441 acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat
     1501 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgcct
     1561 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     1621 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     1681 gccagtgtgc cggtttccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     1741 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     1801 cacagccagt cggtctcagc ttgactttgt cttcacgaag tggctcttca gtggacgaaa
     1861 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1921 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1981 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2041 aaaaaggaag agtatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa
     2101 ataccgctgc gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg
     2161 agaaaatgaa aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga
     2221 tgtggaacgg gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt
     2281 cctgcacttt gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt
     2341 cctttgctcg gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc
     2401 ggagtgcatc aggctctttc actccatcga catatcggat tgtccctata cgaatagctt
     2461 agacagccgc ttagccgaat tggattactt actgaataac gatctggccg atgtggattg
     2521 cgaaaactgg gaagaggaca ctccatttaa agatccgcgc gagctgtatg attttttaaa
     2581 gacggaaaag cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat
     2641 ctttgtgaaa gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga
     2701 caagtggtat gacattgcct tctgcgtccg gtcgctcagg gaggatatcg gggaagaaca
     2761 gtatgtcgag ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata
     2821 ttatatttta ctggatgaat tgttttagct gtcagaccaa gtttactcat atatacttta
     2881 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2941 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3481 aggagagcgc acgagggagc tt
//
