LOCUS       sequence-314361-        6238 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 6238)
  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 6238)
  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_Venus_Hyg
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(98..686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(178..197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(857..1672)
                     /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(857..1672)
                     /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     1580..1599
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1664..1683
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1963..2398
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2525..2547
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    2555..2731
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(2963..3988)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(4008..4353)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(4045..4065)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(5081..5083,5084..5086,5087..5800)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5126..5147)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5387..5406)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5734..5755
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(6221..6238,1..7)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggtaaactaa gtcgctgtat gtgtttgttt gagatctcat gtgagcaaaa ggccagcaaa
       61 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      121 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      181 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      241 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      301 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      361 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      421 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      481 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      541 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaagaaga gttggtagct
      601 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      661 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      721 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      781 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgtgt
      841 aacattggtc tagtgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
      901 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
      961 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1021 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1081 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     1141 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     1201 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     1261 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1321 cacctgaatc aggatattct tctaatacct ggaatgctgt tttccctggg atcgcagtgg
     1381 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     1441 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     1501 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcgg tagattgtcg
     1561 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     1621 tggaatttaa tcgcggcctt gagcaagacg tttcccgttg aatatggctc ataacacccc
     1681 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     1741 gtgcaatgta acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt
     1801 ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa
     1861 agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct
     1921 ccctcacttt ctggctggat gatggggcga ttcaggcgat ccccatccaa cagcccgccg
     1981 tcgagcgggc ttttttatcc ccggaagcct gtggatagag ggtagttatc cacgtgaaac
     2041 cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg gcccgctcca aaaactatcc
     2101 acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg gagtacgtga aatcgctaat
     2161 aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa cgctaatagc cctttcagat
     2221 caacagcttg caaacacccc tcgctccggc aagtagttac agcaagtagt atgttcaatt
     2281 agcttttcaa ttatgaatat atatatcaat tattggtcgc ccttggcttg tggacaatgc
     2341 gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg ttgcccaccg tcgagcgcct
     2401 ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta taaatttttt tttttgaaaa
     2461 agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga tttccgatcc ccggaattag
     2521 atcttggcag gatatattgt ggtgtaacgt tatcgatctg gattttagta ctggattttg
     2581 gttttaggaa ttagaaattt tattgataga agtattttac aaatacaaat acatactaag
     2641 ggtttcttat atgctcaaca catgagcgaa accctataag aaccctaatt tcccttatcg
     2701 ggaaactact cacacattat ttatggagaa aatagagaga gatagatttg tagagagaga
     2761 ctggtgattt cagcgtaccg aattaattcc tgaatcactg cgaccggccc tcccgcgacc
     2821 cagccgagcg agcttagcga actgtggacg agaactgtgc caccaagcgt aaggccgttc
     2881 tctcgcattt gccttgctag gctcgcgcga gttgctggct gaggcgttct cgaaatcagc
     2941 tcttgttcgg tcggcatcta ctctattcct ttgccctcgg acgagtgctg gggcgtcggt
     3001 ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg cttctgcggg
     3061 cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg catcgaccct
     3121 gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt tggtcaagac
     3181 caatgcggag catatacgcc cggagccttg gcgatcctgc aagctccgga tgcctccgct
     3241 cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag aagatgttgg
     3301 cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc
     3361 ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg tgccggactt
     3421 cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg gacgcactga
     3481 cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc gcgcatatga
     3541 aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg aacccgctcg
     3601 tctggctaag atcggccgca gcgatagcat ccatagcctc cgcgaccggc tgaagaacag
     3661 cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc
     3721 aataggtcag gctctcgctg aactccccaa tgtcaagcac ttccggaatc gggagcgcgg
     3781 ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg cagctattta
     3841 cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat tcttcgccct
     3901 ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac ttctcgacag
     3961 acgtcgcggt gagttcaggc tttttcatag gggggatcag cttgggctgt cctctccaaa
     4021 tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca
     4081 tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgaagtcgt ggttggaacg
     4141 acttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca
     4201 gaggcatctt gaatgatagc ctttccttta tcgcaatgat ggcatttgta ggagccacct
     4261 tccttttcta ctgtcctttc gatgaagtga cagatagctg ggcaatggaa tccgaggagg
     4321 tttcccgaaa ttaccctttg ttgaaaagtc tcaataaagc ttcgacgagt cagtaataaa
     4381 cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt ttatcaactc aaagcacaaa
     4441 tacttttcct caacctaaaa ataaggcaat tagccaaaaa caactttgcg tgtaaacaac
     4501 gctcaataca cgtgtcattt tattattagc tattgcttca ccgccttagc tttctcgtga
     4561 cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa caatacccaa agacctcttc
     4621 ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa actttctctc aattctctct
     4681 accgtgatca aggtaaattt ctgtgttcct tattctctca aaatcttcga ttttgttttc
     4741 gttcgatccc aatttcgtat atgttctttg gtttagattc tgttaatctt agatcgaaga
     4801 cgattttctg ggtttgatcg ttagatatca tcttaattct cgattagggt ttcatagata
     4861 tcatccgatt tgttcaaata atttgagttt tgtcgaataa ttactcttcg atttgtgatt
     4921 tctatctaga tctggtgtta gtttctagtt tgtgcgatcg aatttgtcga ttaatctgag
     4981 tttttctgat taacagacta gagttaaccg ggctcaggcc tggcgcgcca ctagtggatc
     5041 catcgatagt actgtcgacc tcgagggtac cgctcccggg atggtgagca agggcgagga
     5101 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     5161 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     5221 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     5281 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     5341 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     5401 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     5461 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     5521 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     5581 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     5641 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     5701 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     5761 cgccgggatc actctcggca tggacgagct gtacaagtaa gagctcatat gaagatgaag
     5821 atgaaatatt tggtgtgtca aataaatagc ttgtgtgctt aagtttgtgt ttttttcttg
     5881 gcttgttgtg ttatgaattt gtggcttttt ctaatattaa atgaatgtaa gatctcatta
     5941 taatgaataa acaaatgttt ctataatcca ttgtgaatgt tttgttggat ctcttctgca
     6001 gcatataact actgtatgtg ctatggtatg gactatggaa tatgattaaa gataaggaat
     6061 tcagatcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg
     6121 cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctcatgatc
     6181 agattgtcgt ttcccgcctt cagtttaaac tatcagtgtt tgacaggata tattggcg
//
