LOCUS       sequence-308937-        7330 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 7330)
  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 7330)
  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_GUS_Hyg
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    625..647
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    655..831
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1063..2088)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(2108..2453)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2145..2165)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             3181..4992
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     misc_feature    5413..5437
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5528..6116)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5608..5627)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6287..7102)
                     /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(6287..7102)
                     /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     7010..7029
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7094..7113
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
       61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
      121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
      181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
      241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
      301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
      361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
      421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
      481 ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta
      541 taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga
      601 tttccgatcc ccggaattag atcttggcag gatatattgt ggtgtaacgt tatcgatctg
      661 gattttagta ctggattttg gttttaggaa ttagaaattt tattgataga agtattttac
      721 aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa accctataag
      781 aaccctaatt tcccttatcg ggaaactact cacacattat ttatggagaa aatagagaga
      841 gatagatttg tagagagaga ctggtgattt cagcgtaccg aattaattcc tgaatcactg
      901 cgaccggccc tcccgcgacc cagccgagcg agcttagcga actgtggacg agaactgtgc
      961 caccaagcgt aaggccgttc tctcgcattt gccttgctag gctcgcgcga gttgctggct
     1021 gaggcgttct cgaaatcagc tcttgttcgg tcggcatcta ctctattcct ttgccctcgg
     1081 acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca
     1141 gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac
     1201 gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct
     1261 ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccttg gcgatcctgc
     1321 aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg
     1381 cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca
     1441 gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc
     1501 gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc
     1561 ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg
     1621 atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc
     1681 gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatagcat ccatagcctc
     1741 cgcgaccggc tgaagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc
     1801 ctgtgcacgg cgggagatgc aataggtcag gctctcgctg aactccccaa tgtcaagcac
     1861 ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa
     1921 accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa
     1981 agcacgagat tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc
     2041 gatcagaaac ttctcgacag acgtcgcggt gagttcaggc tttttcatag gggggatcag
     2101 cttgggctgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag
     2161 gatagtggga ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct
     2221 ttgaagtcgt ggttggaacg acttcttttt ccacgatgct cctcgtgggt gggggtccat
     2281 ctttgggacc actgtcggca gaggcatctt gaatgatagc ctttccttta tcgcaatgat
     2341 ggcatttgta ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg
     2401 ggcaatggaa tccgaggagg tttcccgaaa ttaccctttg ttgaaaagtc tcaataaagc
     2461 ttcgacgagt cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt
     2521 ttatcaactc aaagcacaaa tacttttcct caacctaaaa ataaggcaat tagccaaaaa
     2581 caactttgcg tgtaaacaac gctcaataca cgtgtcattt tattattagc tattgcttca
     2641 ccgccttagc tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa
     2701 caatacccaa agacctcttc ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa
     2761 actttctctc aattctctct accgtgatca aggtaaattt ctgtgttcct tattctctca
     2821 aaatcttcga ttttgttttc gttcgatccc aatttcgtat atgttctttg gtttagattc
     2881 tgttaatctt agatcgaagt cgattttctg ggtttgatcg ttagatatca tcttaattct
     2941 cgattagggt ttcatagata tcatccgatt tgttcaaata atttgagttt tgtcgaataa
     3001 ttactcttcg atttgtgatt tctatctaga tctggtgtta gtttctagtt tgtgcgatcg
     3061 aatttgtcga ttaatctgag tttttctgat taacagacta gagttaaccg ggctcaggcc
     3121 tggcgcgcca ctagtggatc catcgatagt actgtcgacc tcgagggtac cgctcccggg
     3181 atgttacgtc ctgtagaaac cccaacccgt gaaatcaaaa aactcgacgg cctgtgggca
     3241 ttcagtctgg atcgcgaaaa ctgtggaatt gatcagcgtt ggtgggaaag cgcgttacaa
     3301 gaaagccggg caattgctgt gccaggcagt tttaacgatc agttcgccga tgcagatatt
     3361 cgtaattatg cgggcaacgt ctggtatcag cgcgaagtct ttataccgaa aggttgggca
     3421 ggccagcgta tcgtgctgcg tttcgatgcg gtcactcatt acggcaaagt gtgggtcaat
     3481 aatcaggaag tgatggagca tcagggcggc tatacgccat ttgaagccga tgtcacgccg
     3541 tatgttattg ccgggaaaag tgtacgtatc accgtttgtg tgaacaacga actgaactgg
     3601 cagactatcc cgccgggaat ggtgattacc gacgaaaacg gcaagaaaaa gcagtcttac
     3661 ttccatgatt tctttaacta tgccggaatc catcgcagcg taatgctcta caccacgccg
     3721 aacacctggg tggacgatat caccgtggtg acgcatgtcg cgcaagactg taaccacgcg
     3781 tctgttgact ggcaggtggt ggccaatggt gatgtcagcg ttgaactgcg tgatgcggat
     3841 caacaggtgg ttgcaactgg acaaggcact agcgggactt tgcaagtggt gaatccgcac
     3901 ctctggcaac cgggtgaagg ttatctctat gaactgtgcg tcacagccaa aagccagaca
     3961 gagtgtgata tctacccgct tcgcgtcggc atccggtcag tggcagtgaa gggcgaacag
     4021 ttcctgatta accacaaacc gttctacttt actggctttg gtcgtcatga agatgcggac
     4081 ttgcgtggca aaggattcga taacgtgctg atggtgcacg accacgcatt aatggactgg
     4141 attggggcca actcctaccg tacctcgcat tacccttacg ctgaagagat gctcgactgg
     4201 gcagatgaac atggcatcgt ggtgattgat gaaactgctg ctgtcggctt taacctctct
     4261 ttaggcattg gtttcgaagc gggcaacaag ccgaaagaac tgtacagcga agaggcagtc
     4321 aacggggaaa ctcagcaagc gcacttacag gcgattaaag agctgatagc gcgtgacaaa
     4381 aaccacccaa gcgtggtgat gtggagtatt gccaacgaac cggatacccg tccgcaaggt
     4441 gcacgggaat atttcgcgcc actggcggaa gcaacgcgta aactcgaccc gacgcgtccg
     4501 atcacctgcg tcaatgtaat gttctgcgac gctcacaccg ataccatcag cgatctcttt
     4561 gatgtgctgt gcctgaaccg ttattacgga tggtatgtcc aaagcggcga tttggaaacg
     4621 gcagagaagg tactggaaaa agaacttctg gcctggcagg agaaactgca tcagccgatt
     4681 atcatcaccg aatacggcgt ggatacgtta gccgggctgc actcaatgta caccgacatg
     4741 tggagtgaag agtatcagtg tgcatggctg gatatgtatc accgcgtctt tgatcgcgtc
     4801 agcgccgtcg tcggtgaaca ggtatggaat ttcgccgatt ttgcgacctc gcaaggcata
     4861 ttgcgcgttg gcggtaacaa gaaagggatc ttcactcgcg accgcaaacc gaagtcggcg
     4921 gcttttctgc tgcaaaaacg ctggactggc atgaacttcg gtgaaaaacc gcagcaggga
     4981 ggcaaacaat gagagctcat atgaagatga agatgaaata tttggtgtgt caaataaata
     5041 gcttgtgtgc ttaagtttgt gtttttttct tggcttgttg tgttatgaat ttgtggcttt
     5101 ttctaatatt aaatgaatgt aagatctcat tataatgaat aaacaaatgt ttctataatc
     5161 cattgtgaat gttttgttgg atctcttctg cagcatataa ctactgtatg tgctatggta
     5221 tggactatgg aatatgatta aagataagga attcagatcg gctgagtggc tccttcaacg
     5281 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
     5341 aacgtgactc ccttaattct ccgctcatga tcagattgtc gtttcccgcc ttcagtttaa
     5401 actatcagtg tttgacagga tatattggcg ggtaaactaa gtcgctgtat gtgtttgttt
     5461 gagatctcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6001 cagttacctt cggaagaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6241 gttttaaatc aatctaaagt atatatgtgt aacattggtc tagtgattag aaaaactcat
     6301 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     6361 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     6421 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     6481 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     6541 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     6601 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     6661 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     6721 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     6781 ggaatgctgt tttccctggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     6841 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     6901 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     6961 cgggcttccc atacaatcgg tagattgtcg cacctgattg cccgacatta tcgcgagccc
     7021 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctt gagcaagacg
     7081 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     7141 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     7201 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc
     7261 ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca
     7321 cctacaacaa
//
