LOCUS       sequence-71306-e        4114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty destination vector for creating a GreenGate plant 
            transformation plasmid with the plant resistance cassette at the 
            T-DNA left border..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4114)
  AUTHORS   Lampropoulos A, Sutikovic Z, Wenzl C, Maegele I, Lohmann JU, Forner 
            J
  TITLE     GreenGate - A Novel, Versatile, and Efficient Cloning System for 
            Plant Transgenesis.
  JOURNAL   PLoS One. 2013 Dec 20;8(12):e83043. doi: 
            10.1371/journal.pone.0083043.
  PUBMED    24376629
REFERENCE   2  (bases 1 to 4114)
  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: "PLoS One.";
            date: "2013-12-20"; volume: "8(12):e83043. doi"; pages: " 
            10.1371/journal.pone.0083043"
COMMENT     Sequence Label: pGGZ003
FEATURES             Location/Qualifiers
     source          1..4114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    534..556
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             743..1402
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(809..828)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1421..1726
                     /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     1605..1624
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(1756..1778)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1972..1991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    2133..2157
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2248..2836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2328..2347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3007..3795)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(4086..4114,1..407)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 tttttatccc cggaagcctg tggatagagg gtagttatcc acgtgaaacc gctaatgccc
       61 cgcaaagcct tgattcacgg ggctttccgg cccgctccaa aaactatcca cgtgaaatcg
      121 ctaatcaggg tacgtgaaat cgctaatcgg agtacgtgaa atcgctaata aggtcacgtg
      181 aaatcgctaa tcaaaaaggc acgtgagaac gctaatagcc ctttcagatc aacagcttgc
      241 aaacacccct cgctccggca agtagttaca gcaagtagta tgttcaatta gcttttcaat
      301 tatgaatata tatatcaatt attggtcgcc cttggcttgt ggacaatgcg ctacgcgcac
      361 cggctccgcc cgtggacaac cgcaagcggt tgcccaccgt cgagcgcctt tgcccacaac
      421 ccggcggccg gccgcaacag atcgttttat aaattttttt ttttgaaaaa gaaaaagccc
      481 gaaaggcggc aacctctcgg gcttctggat ttccgatccc cggaattaga tcttggcagg
      541 atatattgtg gtgtaacgtt ggatctggct gagaacgcca gctgtgcatg cttggtctag
      601 aatactgaga ccctcgaggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
      661 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggtg
      721 tgctaagctt aggaggtcag ctatggagaa aaaaatcact ggatatacca ccgttgatat
      781 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
      841 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
      901 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggagtt
      961 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1021 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     1081 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1141 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     1201 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1261 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1321 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1381 tgagtggcag ggcggggcgt aagagctcag gaggacagct atgcagttta aggtttacac
     1441 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac
     1501 gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc
     1561 ccgtgaactt tacccagtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     1621 tatggccagt gtgccagtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     1681 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaacta gtaggaggta
     1741 atcaatgctg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1801 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1861 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     1921 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     1981 ctctgatgcc gcatgatcat ctggcggacc tcttcggagg tccgcttttt tttgaattcg
     2041 gtctcaaggt ggtacctggt gcgatcgctg ttggcgcgcc gtgtttaatt aaggttgcgg
     2101 ccgcttactt cgtccgagcc tagttcgagc cttgacagga tatattggcg ggtaaactaa
     2161 gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa
     2221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2701 tatctgcgct ctgctgaagc cagttacctt cggaagaaga gttggtagct cttgatccgg
     2761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgtgt aacattggtc
     3001 tagtgattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgaa caaattcttc
     3061 caactgatct gcgcgcgagg ccaagcgatc ttcttgtcca agataagcct gcctagcttc
     3121 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     3181 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     3241 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     3301 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     3361 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     3421 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     3481 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     3541 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     3601 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     3661 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     3721 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     3781 caccgcttcc ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     3841 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     3901 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     3961 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     4021 caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc
     4081 gatccccatc caacagcccg ccgtcgagcg ggct
//
