LOCUS       sequence-314389-        5539 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 5539)
  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 5539)
  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_6xHis_Hyg
FEATURES             Location/Qualifiers
     source          1..5539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..346)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(38..58)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             1074..1091
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1515..1539
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1630..2218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1710..1729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2389..3204)
                     /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(2389..3204)
                     /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     3112..3131
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3196..3215
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3495..3930
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4057..4079
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    4087..4263
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4495..5520)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 tgtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg
       61 ggattgtgcg tcatccctta cgtcagtgga gatgtcacat caatccactt gctttgaagt
      121 cgtggttgga acgacttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg
      181 accactgtcg gcagaggcat cttgaatgat agcctttcct ttatcgcaat gatggcattt
      241 gtaggagcca ccttcctttt ctactgtcct ttcgatgaag tgacagatag ctgggcaatg
      301 gaatccgagg aggtttcccg aaattaccct ttgttgaaaa gtctcaataa agcttcgacg
      361 agtcagtaat aaacggcgtc aaagtggttg cagccggcac acacgagtcg tgtttatcaa
      421 ctcaaagcac aaatactttt cctcaaccta aaaataaggc aattagccaa aaacaacttt
      481 gcgtgtaaac aacgctcaat acacgtgtca ttttattatt agctattgct tcaccgcctt
      541 agctttctcg tgacctagtc gtcctcgtct tttcttcttc ttcttctata aaacaatacc
      601 caaagacctc ttcttcttca caattcagat ttcaatttct caaaatctta aaaactttct
      661 ctcaattctc tctaccgtga tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt
      721 cgattttgtt ttcgttcgat cccaatttcg tatatgttct ttggtttaga ttctgttaat
      781 cttagatcga agacgatttt ctgggtttga tcgttagata tcatcttaat tctcgattag
      841 ggtttcatag atatcatccg atttgttcaa ataatttgag ttttgtcgaa taattactct
      901 tcgatttgtg atttctatct agatctggtg ttagtttcta gtttgtgcga tcgaatttgt
      961 cgattaatct gagtttttct gattaacaga ctagagttaa ccgggctcag gcctggcgcg
     1021 ccactagtgg atccatcgat agtactgtcg acctcgaggg taccgctccc gggcatcacc
     1081 atcaccatca ctaagagctc atatgaagat gaagatgaaa tatttggtgt gtcaaataaa
     1141 tagcttgtgt gcttaagttt gtgttttttt cttggcttgt tgtgttatga atttgtggct
     1201 ttttctaata ttaaatgaat gtaagatctc attataatga ataaacaaat gtttctataa
     1261 tccattgtga atgttttgtt ggatctcttc tgcagcatat aactactgta tgtgctatgg
     1321 tatggactat ggaatatgat taaagataag gaattcagat cggctgagtg gctccttcaa
     1381 cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc
     1441 ataacgtgac tcccttaatt ctccgctcat gatcagattg tcgtttcccg ccttcagttt
     1501 aaactatcag tgtttgacag gatatattgg cgggtaaact aagtcgctgt atgtgtttgt
     1561 ttgagatctc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2041 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2101 gccagttacc ttcggaagaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2341 aagttttaaa tcaatctaaa gtatatatgt gtaacattgg tctagtgatt agaaaaactc
     2401 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     2461 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     2521 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     2581 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     2641 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     2701 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     2761 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     2821 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     2881 ctggaatgct gttttccctg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     2941 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     3001 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     3061 atcgggcttc ccatacaatc ggtagattgt cgcacctgat tgcccgacat tatcgcgagc
     3121 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc ttgagcaaga
     3181 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     3241 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     3301 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agatcacgca
     3361 tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc
     3421 cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg atgatggggc
     3481 gattcaggcg atccccatcc aacagcccgc cgtcgagcgg gcttttttat ccccggaagc
     3541 ctgtggatag agggtagtta tccacgtgaa accgctaatg ccccgcaaag ccttgattca
     3601 cggggctttc cggcccgctc caaaaactat ccacgtgaaa tcgctaatca gggtacgtga
     3661 aatcgctaat cggagtacgt gaaatcgcta ataaggtcac gtgaaatcgc taatcaaaaa
     3721 ggcacgtgag aacgctaata gccctttcag atcaacagct tgcaaacacc cctcgctccg
     3781 gcaagtagtt acagcaagta gtatgttcaa ttagcttttc aattatgaat atatatatca
     3841 attattggtc gcccttggct tgtggacaat gcgctacgcg caccggctcc gcccgtggac
     3901 aaccgcaagc ggttgcccac cgtcgagcgc ctttgcccac aacccggcgg ccggccgcaa
     3961 cagatcgttt tataaatttt tttttttgaa aaagaaaaag cccgaaaggc ggcaacctct
     4021 cgggcttctg gatttccgat ccccggaatt agatcttggc aggatatatt gtggtgtaac
     4081 gttatcgatc tggattttag tactggattt tggttttagg aattagaaat tttattgata
     4141 gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg
     4201 aaaccctata agaaccctaa tttcccttat cgggaaacta ctcacacatt atttatggag
     4261 aaaatagaga gagatagatt tgtagagaga gactggtgat ttcagcgtac cgaattaatt
     4321 cctgaatcac tgcgaccggc cctcccgcga cccagccgag cgagcttagc gaactgtgga
     4381 cgagaactgt gccaccaagc gtaaggccgt tctctcgcat ttgccttgct aggctcgcgc
     4441 gagttgctgg ctgaggcgtt ctcgaaatca gctcttgttc ggtcggcatc tactctattc
     4501 ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca
     4561 gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc
     4621 tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc
     4681 gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagcct
     4741 tggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca
     4801 agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat
     4861 cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga
     4921 gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag
     4981 ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg
     5041 atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat
     5101 tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatagc
     5161 atccatagcc tccgcgaccg gctgaagaac agcgggcagt tcggtttcag gcaggtcttg
     5221 caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaactcccc
     5281 aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg
     5341 atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac
     5401 atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct
     5461 gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat
     5521 aggggggatc agcttgggc
//
