LOCUS       sequence-322859-        6415 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 6415)
  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 6415)
  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_Blp
FEATURES             Location/Qualifiers
     source          1..6415
                     /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..1103
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1527..1551
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1642..2230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1722..1741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3739..4554)
                     /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(3739..4554)
                     /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     4462..4481
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4546..4565
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4845..5280
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5407..5429
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    5437..5613
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(5845..6396)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
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 ggggagcaga
     1081 agctgatctc ggaggaagac ttgtaagagc tcatatgaag atgaagatga aatatttggt
     1141 gtgtcaaata aatagcttgt gtgcttaagt ttgtgttttt ttcttggctt gttgtgttat
     1201 gaatttgtgg ctttttctaa tattaaatga atgtaagatc tcattataat gaataaacaa
     1261 atgtttctat aatccattgt gaatgttttg ttggatctct tctgcagcat ataactactg
     1321 tatgtgctat ggtatggact atggaatatg attaaagata aggaattcag atcggctgag
     1381 tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca
     1441 tcggcggggg tcataacgtg actcccttaa ttctccgctc atgatcagat tgtcgtttcc
     1501 cgccttcagt ttaaactatc agtgtttgac aggatatatt ggcgggtaaa ctaagtcgct
     1561 gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2041 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2101 cgctctgctg aagccagtta ccttcggaag aagagttggt agctcttgat ccggcaaaca
     2161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2341 aaattaaaaa tgaagtttta aatcaatcta aagtactgat gaatccccta atgattttta
     2401 tcaaaatcat taagttaagg tagatacaca tcttgtcata tgatcaaatg gtttcgccaa
     2461 aaatcaataa tcagacaaca aaatgtgcga actcgatatt ttacacgact ctctttacca
     2521 attctgcccc gaattacact taaaacgact caacagctta acgttggctt gccacgcctt
     2581 acttgactgt aaaactctca ctcttaccga acttggccgt aacctgccaa ccaaagcgag
     2641 aacaaaacat aacatcaaac gaatcgaccg attgttaggt aatcgtcacc tccacaaaga
     2701 gcgactcgct gtataccgtt ggcatgctag ctttatctgt tcgggcaata cgatgcccat
     2761 tgtacttgtt gactggtctg atatccgtga gcaaaaacgg cttatggtat tgcgagcttc
     2821 agtcgcacta cacggtcgtt ctgttactct ttatgagaaa gcgttcccgc tttcagagca
     2881 atgttcaaag aaagctcatg accaatttct agccgacctt gcgagcattc taccgagtaa
     2941 caccacaccg ctcattgtca gtgatgctgg ctttaaagtg ccatggtata aatccgttga
     3001 gaagctgggt tggtactggt taagtcgagt aagaggaaaa gtacaatatg cagacctagg
     3061 agcggaaaac tggaaaccta tcagcaactt acatgatatg tcatctagtc actcaaagac
     3121 tttaggctat aagaggctga ctaaaagcaa tccaatctca tgccaaattc tattgtataa
     3181 atctcgctct aaaggccgaa aaaatcagcg ctcgacacgg actcattgtc accacccgtc
     3241 acctaaaatc tactcagcgt cggcaaagga gccatggatt ctagcaacta acttacctgt
     3301 tgaaattcga acacccaaac aacttgttaa tatctattcg aagcgaatgc agattgaaga
     3361 aaccttccga gacttgaaaa gtcctgccta cggactaggc ctacgccata gccgaacgag
     3421 cagctcagag cgttttgata tcatgctgct aatcgccctg atgcttcaac taacatgttg
     3481 gcttgcgggc gttcatgctc agaaacaagg ttgggacaag cacttccagg ctaacacagt
     3541 cagaaatcga aacgtactct caacagttcg cttaggcatg gaagttttgc ggcattctgg
     3601 ctacacaata acaagggaag acttactcgt ggctgcaacc ctactagctc aaaatttatt
     3661 cacacatggt tacgctttgg ggaaattatg aggggatctc tcagtctaaa gtatatatgt
     3721 gtaacattgg tctagtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3781 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3841 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3901 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3961 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     4021 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     4081 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     4141 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     4201 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt
     4261 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     4321 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4381 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt
     4441 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4501 gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacacc
     4561 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     4621 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     4681 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     4741 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     4801 ctccctcact ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc
     4861 cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa
     4921 accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat
     4981 ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta
     5041 ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag
     5101 atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa
     5161 ttagcttttc aattatgaat atatatatca attattggtc gcccttggct tgtggacaat
     5221 gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc
     5281 ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa
     5341 aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt
     5401 agatcttggc aggatatatt gtggtgtaac gttatcgatc tggattttag tactggattt
     5461 tggttttagg aattagaaat tttattgata gaagtatttt acaaatacaa atacatacta
     5521 agggtttctt atatgctcaa cacatgagcg aaaccctata agaaccctaa tttcccttat
     5581 cgggaaacta ctcacacatt atttatggag aaaatagaga gagatagatt tgtagagaga
     5641 gactggtgat ttcagcgtac cgaattaatt cctgaatcac tgcgaccggc cctcccgcga
     5701 cccagccgag cgagcttagc gaactgtgga cgagaactgt gccaccaagc gtaaggccgt
     5761 tctctcgcat ttgccttgct aggctcgcgc gagttgctgg ctgaggcgtt ctcgaaatca
     5821 gctcttgttc ggtcggcatc tacttcagat ttcggtgacg ggcaggaccg gacggggcgg
     5881 caccggcagg ctgaagtcca gctgccagaa acccacgtca tgccagttcc cgtgcttgaa
     5941 gccggccgcc cgcagcatgc cgcggggggc atatccgagc gcctcgtgca tgcgcacgct
     6001 cgggtcgttg ggcagcccga tgacagcgac cacgctcttg aagccctgtg cctccaggga
     6061 cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc ggggggagac
     6121 gtacacggtg gactcggccg tccagtcgta ggcgttgcgt gccttccagg gacccgcgta
     6181 ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc cagggatagc gctcccgcag
     6241 acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga agttgaccgt
     6301 gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg cctcggtggc
     6361 acggcggatg tcggccgggc gtcgttctgg gctcataggg gggatcagct tgggc
//
