LOCUS       sequence-333788-        5551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGREEN-based cloning construct for expression in planta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5551)
  AUTHORS   Pratt AI, Knoblauch J, Kunz HH
  TITLE     An updated pGREEN-based vector suite for cost-effective cloning in 
            plant molecular biology.
  JOURNAL   MicroPubl Biol. 2020 Oct 7;2020. doi: 
            10.17912/micropub.biology.000317.
  PUBMED    33274321
REFERENCE   2  (bases 1 to 5551)
  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: "MicroPubl 
            Biol."; date: "2020-10-7"; volume: "2020. doi"; pages: " 
            10.17912/micropub.biology.000317"
COMMENT     Sequence Label: pG20_Myc_Hyg
FEATURES             Location/Qualifiers
     source          1..5551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(88..676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(168..187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(847..1662)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(847..1662)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1570..1589
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1654..1673
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1953..2388
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2515..2537
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    2545..2721
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(2953..3978)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(3998..4343)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(4035..4055)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5071..5100
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5524..5548
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa
       61 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      541 tatctgcgct ctgctgaagc cagttacctt cggaagaaga gttggtagct cttgatccgg
      601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgtgt aacattggtc
      841 tagtgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
      901 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
      961 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     1021 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     1081 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     1141 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     1201 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     1261 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     1321 aggatattct tctaatacct ggaatgctgt tttccctggg atcgcagtgg tgagtaacca
     1381 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     1441 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     1501 cagaaacaac tctggcgcat cgggcttccc atacaatcgg tagattgtcg cacctgattg
     1561 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     1621 tcgcggcctt gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     1681 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     1741 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     1801 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     1861 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt
     1921 ctggctggat gatggggcga ttcaggcgat ccccatccaa cagcccgccg tcgagcgggc
     1981 ttttttatcc ccggaagcct gtggatagag ggtagttatc cacgtgaaac cgctaatgcc
     2041 ccgcaaagcc ttgattcacg gggctttccg gcccgctcca aaaactatcc acgtgaaatc
     2101 gctaatcagg gtacgtgaaa tcgctaatcg gagtacgtga aatcgctaat aaggtcacgt
     2161 gaaatcgcta atcaaaaagg cacgtgagaa cgctaatagc cctttcagat caacagcttg
     2221 caaacacccc tcgctccggc aagtagttac agcaagtagt atgttcaatt agcttttcaa
     2281 ttatgaatat atatatcaat tattggtcgc ccttggcttg tggacaatgc gctacgcgca
     2341 ccggctccgc ccgtggacaa ccgcaagcgg ttgcccaccg tcgagcgcct ttgcccacaa
     2401 cccggcggcc ggccgcaaca gatcgtttta taaatttttt tttttgaaaa agaaaaagcc
     2461 cgaaaggcgg caacctctcg ggcttctgga tttccgatcc ccggaattag atcttggcag
     2521 gatatattgt ggtgtaacgt tatcgatctg gattttagta ctggattttg gttttaggaa
     2581 ttagaaattt tattgataga agtattttac aaatacaaat acatactaag ggtttcttat
     2641 atgctcaaca catgagcgaa accctataag aaccctaatt tcccttatcg ggaaactact
     2701 cacacattat ttatggagaa aatagagaga gatagatttg tagagagaga ctggtgattt
     2761 cagcgtaccg aattaattcc tgaatcactg cgaccggccc tcccgcgacc cagccgagcg
     2821 agcttagcga actgtggacg agaactgtgc caccaagcgt aaggccgttc tctcgcattt
     2881 gccttgctag gctcgcgcga gttgctggct gaggcgttct cgaaatcagc tcttgttcgg
     2941 tcggcatcta ctctattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc
     3001 ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt
     3061 acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc
     3121 tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag
     3181 catatacgcc cggagccttg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg
     3241 cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta
     3301 ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc
     3361 cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc
     3421 ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc
     3481 catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca
     3541 tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag
     3601 atcggccgca gcgatagcat ccatagcctc cgcgaccggc tgaagaacag cgggcagttc
     3661 ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag
     3721 gctctcgctg aactccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa
     3781 gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac
     3841 atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg
     3901 catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt
     3961 gagttcaggc tttttcatag gggggatcag cttgggctgt cctctccaaa tgaaatgaac
     4021 ttccttatat agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt
     4081 cagtggagat gtcacatcaa tccacttgct ttgaagtcgt ggttggaacg acttcttttt
     4141 ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca gaggcatctt
     4201 gaatgatagc ctttccttta tcgcaatgat ggcatttgta ggagccacct tccttttcta
     4261 ctgtcctttc gatgaagtga cagatagctg ggcaatggaa tccgaggagg tttcccgaaa
     4321 ttaccctttg ttgaaaagtc tcaataaagc ttcgacgagt cagtaataaa cggcgtcaaa
     4381 gtggttgcag ccggcacaca cgagtcgtgt ttatcaactc aaagcacaaa tacttttcct
     4441 caacctaaaa ataaggcaat tagccaaaaa caactttgcg tgtaaacaac gctcaataca
     4501 cgtgtcattt tattattagc tattgcttca ccgccttagc tttctcgtga cctagtcgtc
     4561 ctcgtctttt cttcttcttc ttctataaaa caatacccaa agacctcttc ttcttcacaa
     4621 ttcagatttc aatttctcaa aatcttaaaa actttctctc aattctctct accgtgatca
     4681 aggtaaattt ctgtgttcct tattctctca aaatcttcga ttttgttttc gttcgatccc
     4741 aatttcgtat atgttctttg gtttagattc tgttaatctt agatcgaaga cgattttctg
     4801 ggtttgatcg ttagatatca tcttaattct cgattagggt ttcatagata tcatccgatt
     4861 tgttcaaata atttgagttt tgtcgaataa ttactcttcg atttgtgatt tctatctaga
     4921 tctggtgtta gtttctagtt tgtgcgatcg aatttgtcga ttaatctgag tttttctgat
     4981 taacagacta gagttaaccg ggctcaggcc tggcgcgcca ctagtggatc catcgatagt
     5041 actgtcgacc tcgagggtac cgctcccggg gagcagaagc tgatctcgga ggaagacttg
     5101 taagagctca tatgaagatg aagatgaaat atttggtgtg tcaaataaat agcttgtgtg
     5161 cttaagtttg tgtttttttc ttggcttgtt gtgttatgaa tttgtggctt tttctaatat
     5221 taaatgaatg taagatctca ttataatgaa taaacaaatg tttctataat ccattgtgaa
     5281 tgttttgttg gatctcttct gcagcatata actactgtat gtgctatggt atggactatg
     5341 gaatatgatt aaagataagg aattcagatc ggctgagtgg ctccttcaac gttgcggttc
     5401 tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact
     5461 cccttaattc tccgctcatg atcagattgt cgtttcccgc cttcagttta aactatcagt
     5521 gtttgacagg atatattggc gggtaaacta a
//
