LOCUS       sequence-319741-        3497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate sgRNA array cloning vector
            1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3497)
  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 3497)
  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: pDGE503
FEATURES             Location/Qualifiers
     source          1..3497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..132
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        222..324
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             325..1008
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(391..410)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1328..1633
                     /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     1512..1531
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(1714..1732)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1800..1904
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1905..2699
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      2870..3458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3359..3378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
       61 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      121 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
      181 ccgattcatt aatcactctg tgaagacatt tgctgagacc ttgatcggca cgtaagaggt
      241 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
      301 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
      361 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
      421 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
      481 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
      541 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
      601 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
      661 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
      721 tatttcccta aagggtttat tgagaatatg tttttcgttt cagccaatcc ctgggtgagt
      781 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
      841 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
      901 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
      961 gatgagtggc agggggggcg taaacgccgc gtggatccgg cttactaaaa gccagataac
     1021 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     1081 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     1141 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     1201 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     1261 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     1321 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     1381 ggatgtacag agtgatatta ttgacacgcc tgggcgacgg atggtgatcc ccctggccag
     1441 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     1501 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtttccg ttatcgggga
     1561 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     1621 ctggggaata taaatgtcag gctcccttat acacagccag tcggtctcag cttccctttg
     1681 tcttcacgaa gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg
     1741 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1801 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1861 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatggct aaaatgagaa
     1921 tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg cgtaaaagat acggaaggaa
     1981 tgtctcctgc taaggtatat aagctggtgg gagaaaatga aaacctatat ttaaaaatga
     2041 cggacagccg gtataaaggg accacctatg atgtggaacg ggaaaaggac atgatgctat
     2101 ggctggaagg aaagctgcct gttccaaagg tcctgcactt tgaacggcat gatggctgga
     2161 gcaatctgct catgagtgag gccgatggcg tcctttgctc ggaagagtat gaagatgaac
     2221 aaagccctga aaagattatc gagctgtatg cggagtgcat caggctcttt cactccatcg
     2281 acatatcgga ttgtccctat acgaatagct tagacagccg cttagccgaa ttggattact
     2341 tactgaataa cgatctggcc gatgtggatt gcgaaaactg ggaagaggac actccattta
     2401 aagatccgcg cgagctgtat gattttttaa agacggaaaa gcccgaagag gaacttgtct
     2461 tttcccacgg cgacctggga gacagcaaca tctttgtgaa agatggcaaa gtaagtggct
     2521 ttattgatct tgggagaagc ggcagggcgg acaagtggta tgacattgcc ttctgcgtcc
     2581 ggtcgctcag ggaggatatc ggggaagaac agtatgtcga gctatttttt gacttactgg
     2641 ggatcaagcc tgattgggag aaaataaaat attatatttt actggatgaa ttgttttagc
     2701 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2761 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2821 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2881 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2941 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3001 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3061 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3121 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3181 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3241 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3301 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3361 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3421 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3481 tacggttcct ggccttt
//
