LOCUS       sequence-314406-        8668 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 8668)
  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 8668)
  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..8668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             728..2539
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTGCDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     misc_feature    2960..2984
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3075..3663)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3155..3174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5172..5987)
                     /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(5172..5987)
                     /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     5895..5914
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5979..5998
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6278..6713
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    6840..6862
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    6870..7046
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7278..8303)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(8323..8668)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(8360..8380)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 ataaagcttc gacgagtcag taataaacgg cgtcaaagtg gttgcagccg gcacacacga
       61 gtcgtgttta tcaactcaaa gcacaaatac ttttcctcaa cctaaaaata aggcaattag
      121 ccaaaaacaa ctttgcgtgt aaacaacgct caatacacgt gtcattttat tattagctat
      181 tgcttcaccg ccttagcttt ctcgtgacct agtcgtcctc gtcttttctt cttcttcttc
      241 tataaaacaa tacccaaaga cctcttcttc ttcacaattc agatttcaat ttctcaaaat
      301 cttaaaaact ttctctcaat tctctctacc gtgatcaagg taaatttctg tgttccttat
      361 tctctcaaaa tcttcgattt tgttttcgtt cgatcccaat ttcgtatatg ttctttggtt
      421 tagattctgt taatcttaga tcgaagacga ttttctgggt ttgatcgtta gatatcatct
      481 taattctcga ttagggtttc atagatatca tccgatttgt tcaaataatt tgagttttgt
      541 cgaataatta ctcttcgatt tgtgatttct atctagatct ggtgttagtt tctagtttgt
      601 gcgatcgaat ttgtcgatta atctgagttt ttctgattaa cagactagag ttaaccgggc
      661 tcaggcctgg cgcgccacta gtggatccat cgatagtact gtcgacctcg agggtaccgc
      721 tcccgggatg ttacgtcctg tagaaacccc aacccgtgaa atcaaaaaac tcgacggcct
      781 gtgggcattc agtctggatc gcgaaaactg tggaattgat cagcgttggt gggaaagcgc
      841 gttacaagaa agccgggcaa ttgctgtgcc aggcagtttt aacgatcagt tcgccgatgc
      901 agatattcgt aattatgcgg gcaacgtctg gtatcagcgc gaagtcttta taccgaaagg
      961 ttgggcaggc cagcgtatcg tgctgcgttt cgatgcggtc actcattacg gcaaagtgtg
     1021 ggtcaataat caggaagtga tggagcatca gggcggctat acgccatttg aagccgatgt
     1081 cacgccgtat gttattgccg ggaaaagtgt acgtatcacc gtttgtgtga acaacgaact
     1141 gaactggcag actatcccgc cgggaatggt gattaccgac gaaaacggca agaaaaagca
     1201 gtcttacttc catgatttct ttaactatgc cggaatccat cgcagcgtaa tgctctacac
     1261 cacgccgaac acctgggtgg acgatatcac cgtggtgacg catgtcgcgc aagactgtaa
     1321 ccacgcgtct gttgactggc aggtggtggc caatggtgat gtcagcgttg aactgcgtga
     1381 tgcggatcaa caggtggttg caactggaca aggcactagc gggactttgc aagtggtgaa
     1441 tccgcacctc tggcaaccgg gtgaaggtta tctctatgaa ctgtgcgtca cagccaaaag
     1501 ccagacaggg tgtgatatct acccgcttcg cgtcggcatc cggtcagtgg cagtgaaggg
     1561 cgaacagttc ctgattaacc acaaaccgtt ctactttact ggctttggtc gtcatgaaga
     1621 tgcggacttg cgtggcaaag gattcgataa cgtgctgatg gtgcacgacc acgcattaat
     1681 ggactggatt ggggccaact cctaccgtac ctcgcattac ccttacgctg aagagatgct
     1741 cgactgggca gatgaacatg gcatcgtggt gattgatgaa actgctgctg tcggctttaa
     1801 cctctcttta ggcattggtt tcgaagcggg caacaagccg aaagaactgt acagcgaaga
     1861 ggcagtcaac ggggaaactc agcaagcgca cttacaggcg attaaagagc tgatagcgcg
     1921 tgacaaaaac cacccaagcg tggtgatgtg gagtattgcc aacgaaccgg atacccgtcc
     1981 gcaaggtgca cgggaatatt tcgcgccact ggcggaagca acgcgtaaac tcgacccgac
     2041 gcgtccgatc acctgcgtca atgtaatgtt ctgcgacgct cacaccgata ccatcagcga
     2101 tctctttgat gtgctgtgcc tgaaccgtta ttacggatgg tatgtccaaa gcggcgattt
     2161 ggaaacggca gagaaggtac tggaaaaaga acttctggcc tggcaggaga aactgcatca
     2221 gccgattatc atcaccgaat acggcgtgga tacgttagcc gggctgcact caatgtacac
     2281 cgacatgtgg agtgaagagt atcagtgtgc atggctggat atgtatcacc gcgtctttga
     2341 tcgcgtcagc gccgtcgtcg gtgaacaggt atggaatttc gccgattttg cgacctcgca
     2401 aggcatattg cgcgttggcg gtaacaagaa agggatcttc actcgcgacc gcaaaccgaa
     2461 gtcggcggct tttctgctgc aaaaacgctg gactggcatg aacttcggtg aaaaaccgca
     2521 gcagggaggc aaacaatgag agctcatatg aagatgaaga tgaaatattt ggtgtgtcaa
     2581 ataaatagct tgtgtgctta agtttgtgtt tttttcttgg cttgttgtgt tatgaatttg
     2641 tggctttttc taatattaaa tgaatgtaag atctcattat aatgaataaa caaatgtttc
     2701 tataatccat tgtgaatgtt ttgttggatc tcttctgcag catataacta ctgtatgtgc
     2761 tatggtatgg actatggaat atgattaaag ataaggaatt cagatcggct gagtggctcc
     2821 ttcaacgttg cggttctgtc agttccaaac gtaaaacggc ttgtcccgcg tcatcggcgg
     2881 gggtcataac gtgactccct taattctccg ctcatgatca gattgtcgtt tcccgccttc
     2941 agtttaaact atcagtgttt gacaggatat attggcgggt aaactaagtc gctgtatgtg
     3001 tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3061 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3121 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3181 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3241 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3301 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3361 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3421 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3481 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3541 ctgaagccag ttaccttcgg aagaagagtt ggtagctctt gatccggcaa acaaaccacc
     3601 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3661 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3721 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3781 aaatgaagtt ttaaatcaat ctaaagtact gatgaatccc ctaatgattt ttatcaaaat
     3841 cattaagtta aggtagatac acatcttgtc atatgatcaa atggtttcgc caaaaatcaa
     3901 taatcagaca acaaaatgtg cgaactcgat attttacacg actctcttta ccaattctgc
     3961 cccgaattac acttaaaacg actcaacagc ttaacgttgg cttgccacgc cttacttgac
     4021 tgtaaaactc tcactcttac cgaacttggc cgtaacctgc caaccaaagc gagaacaaaa
     4081 cataacatca aacgaatcga ccgattgtta ggtaatcgtc acctccacaa agagcgactc
     4141 gctgtatacc gttggcatgc tagctttatc tgttcgggca atacgatgcc cattgtactt
     4201 gttgactggt ctgatatccg tgagcaaaaa cggcttatgg tattgcgagc ttcagtcgca
     4261 ctacacggtc gttctgttac tctttatgag aaagcgttcc cgctttcaga gcaatgttca
     4321 aagaaagctc atgaccaatt tctagccgac cttgcgagca ttctaccgag taacaccaca
     4381 ccgctcattg tcagtgatgc tggctttaaa gtgccatggt ataaatccgt tgagaagctg
     4441 ggttggtact ggttaagtcg agtaagagga aaagtacaat atgcagacct aggagcggaa
     4501 aactggaaac ctatcagcaa cttacatgat atgtcatcta gtcactcaaa gactttaggc
     4561 tataagaggc tgactaaaag caatccaatc tcatgccaaa ttctattgta taaatctcgc
     4621 tctaaaggcc gaaaaaatca gcgctcgaca cggactcatt gtcaccaccc gtcacctaaa
     4681 atctactcag cgtcggcaaa ggagccatgg attctagcaa ctaacttacc tgttgaaatt
     4741 cgaacaccca aacaacttgt taatatctat tcgaagcgaa tgcagattga agaaaccttc
     4801 cgagacttga aaagtcctgc ctacggacta ggcctacgcc atagccgaac gagcagctca
     4861 gagcgttttg atatcatgct gctaatcgcc ctgatgcttc aactaacatg ttggcttgcg
     4921 ggcgttcatg ctcagaaaca aggttgggac aagcacttcc aggctaacac agtcagaaat
     4981 cgaaacgtac tctcaacagt tcgcttaggc atggaagttt tgcggcattc tggctacaca
     5041 ataacaaggg aagacttact cgtggctgca accctactag ctcaaaattt attcacacat
     5101 ggttacgctt tggggaaatt atgaggggat ctctcagtct aaagtatata tgtgtaacat
     5161 tggtctagtg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     5221 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     5281 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     5341 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     5401 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     5461 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     5521 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     5581 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     5641 gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc agtggtgagt
     5701 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     5761 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     5821 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat tgtcgcacct
     5881 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     5941 tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcataac accccttgta
     6001 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     6061 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     6121 gagttgaagg atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa
     6181 aagttcaaaa tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc
     6241 actttctggc tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag
     6301 cgggcttttt tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta
     6361 atgccccgca aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg
     6421 aaatcgctaa tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt
     6481 cacgtgaaat cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca
     6541 gcttgcaaac acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt
     6601 ttcaattatg aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac
     6661 gcgcaccggc tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgcctttgcc
     6721 cacaacccgg cggccggccg caacagatcg ttttataaat tttttttttt gaaaaagaaa
     6781 aagcccgaaa ggcggcaacc tctcgggctt ctggatttcc gatccccgga attagatctt
     6841 ggcaggatat attgtggtgt aacgttatcg atctggattt tagtactgga ttttggtttt
     6901 aggaattaga aattttattg atagaagtat tttacaaata caaatacata ctaagggttt
     6961 cttatatgct caacacatga gcgaaaccct ataagaaccc taatttccct tatcgggaaa
     7021 ctactcacac attatttatg gagaaaatag agagagatag atttgtagag agagactggt
     7081 gatttcagcg taccgaatta attcctgaat cactgcgacc ggccctcccg cgacccagcc
     7141 gagcgagctt agcgaactgt ggacgagaac tgtgccacca agcgtaaggc cgttctctcg
     7201 catttgcctt gctaggctcg cgcgagttgc tggctgaggc gttctcgaaa tcagctcttg
     7261 ttcggtcggc atctactcta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca
     7321 ctatcggcga gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt
     7381 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc
     7441 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg
     7501 cggagcatat acgcccggag ccttggcgat cctgcaagct ccggatgcct ccgctcgaag
     7561 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc
     7621 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca
     7681 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg
     7741 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg
     7801 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca
     7861 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg
     7921 ctaagatcgg ccgcagcgat agcatccata gcctccgcga ccggctgaag aacagcgggc
     7981 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag
     8041 gtcaggctct cgctgaactc cccaatgtca agcacttccg gaatcgggag cgcggccgat
     8101 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc
     8161 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag
     8221 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc
     8281 gcggtgagtt caggcttttt catagggggg atcagcttgg gctgtcctct ccaaatgaaa
     8341 tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct
     8401 tacgtcagtg gagatgtcac atcaatccac ttgctttgaa gtcgtggttg gaacgacttc
     8461 tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagaggc
     8521 atcttgaatg atagcctttc ctttatcgca atgatggcat ttgtaggagc caccttcctt
     8581 ttctactgtc ctttcgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc
     8641 cgaaattacc ctttgttgaa aagtctca
//
