LOCUS       sequence-314457-        5002 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 5002)
  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 5002)
  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_MCS_blp
FEATURES             Location/Qualifiers
     source          1..5002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(100..688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(180..199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(859..1674)
                     /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(859..1674)
                     /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     1582..1601
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1666..1685
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1965..2400
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    2527..2549
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    2557..2733
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(2965..3516)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(3536..3881)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3573..3593)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    join(4987..5002,1..9)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca
       61 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa gagttggtag
      601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatgt
      841 gtaacattgg tctagtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      901 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      961 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1021 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1081 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1141 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1201 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     1261 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1321 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt
     1381 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     1441 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1501 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt
     1561 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1621 gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacacc
     1681 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     1741 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     1801 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     1861 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     1921 ctccctcact ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc
     1981 cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa
     2041 accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat
     2101 ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta
     2161 ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag
     2221 atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa
     2281 ttagcttttc aattatgaat atatatatca attattggtc gcccttggct tgtggacaat
     2341 gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc
     2401 ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa
     2461 aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt
     2521 agatcttggc aggatatatt gtggtgtaac gttatcgatc tggattttag tactggattt
     2581 tggttttagg aattagaaat tttattgata gaagtatttt acaaatacaa atacatacta
     2641 agggtttctt atatgctcaa cacatgagcg aaaccctata agaaccctaa tttcccttat
     2701 cgggaaacta ctcacacatt atttatggag aaaatagaga gagatagatt tgtagagaga
     2761 gactggtgat ttcagcgtac cgaattaatt cctgaatcac tgcgaccggc cctcccgcga
     2821 cccagccgag cgagcttagc gaactgtgga cgagaactgt gccaccaagc gtaaggccgt
     2881 tctctcgcat ttgccttgct aggctcgcgc gagttgctgg ctgaggcgtt ctcgaaatca
     2941 gctcttgttc ggtcggcatc tacttcagat ttcggtgacg ggcaggaccg gacggggcgg
     3001 caccggcagg ctgaagtcca gctgccagaa acccacgtca tgccagttcc cgtgcttgaa
     3061 gccggccgcc cgcagcatgc cgcggggggc atatccgagc gcctcgtgca tgcgcacgct
     3121 cgggtcgttg ggcagcccga tgacagcgac cacgctcttg aagccctgtg cctccaggga
     3181 cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc ggggggagac
     3241 gtacacggtg gactcggccg tccagtcgta ggcgttgcgt gccttccagg gacccgcgta
     3301 ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc cagggatagc gctcccgcag
     3361 acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga agttgaccgt
     3421 gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg cctcggtggc
     3481 acggcggatg tcggccgggc gtcgttctgg gctcataggg gggatcagct tgggctgtcc
     3541 tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga tagtgggatt
     3601 gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc cacttgcttt gaagtcgtgg
     3661 ttggaacgac ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac
     3721 tgtcggcaga ggcatcttga atgatagcct ttcctttatc gcaatgatgg catttgtagg
     3781 agccaccttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc
     3841 cgaggaggtt tcccgaaatt accctttgtt gaaaagtctc aataaagctt cgacgagtca
     3901 gtaataaacg gcgtcaaagt ggttgcagcc ggcacacacg agtcgtgttt atcaactcaa
     3961 agcacaaata cttttcctca acctaaaaat aaggcaatta gccaaaaaca actttgcgtg
     4021 taaacaacgc tcaatacacg tgtcatttta ttattagcta ttgcttcacc gccttagctt
     4081 tctcgtgacc tagtcgtcct cgtcttttct tcttcttctt ctataaaaca atacccaaag
     4141 acctcttctt cttcacaatt cagatttcaa tttctcaaaa tcttaaaaac tttctctcaa
     4201 ttctctctac cgtgatcaag gtaaatttct gtgttcctta ttctctcaaa atcttcgatt
     4261 ttgttttcgt tcgatcccaa tttcgtatat gttctttggt ttagattctg ttaatcttag
     4321 atcgaagacg attttctggg tttgatcgtt agatatcatc ttaattctcg attagggttt
     4381 catagatatc atccgatttg ttcaaataat ttgagttttg tcgaataatt actcttcgat
     4441 ttgtgatttc tatctagatc tggtgttagt ttctagtttg tgcgatcgaa tttgtcgatt
     4501 aatctgagtt tttctgatta acagactagt ggatccgtcg acctcgaggg taccgctccc
     4561 gggtaggagc tcatatgaag atgaagatga aatatttggt gtgtcaaata aatagcttgt
     4621 gtgcttaagt ttgtgttttt ttcttggctt gttgtgttat gaatttgtgg ctttttctaa
     4681 tattaaatga atgtaagatc tcattataat gaataaacaa atgtttctat aatccattgt
     4741 gaatgttttg ttggatctct tctgcagcat ataactactg tatgtgctat ggtatggact
     4801 atggaatatg attaaagata aggaattcag atcggctgag tggctccttc aacgttgcgg
     4861 ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca tcggcggggg tcataacgtg
     4921 actcccttaa ttctccgctc atgatcagat tgtcgtttcc cgccttcagt ttaaactatc
     4981 agtgtttgac aggatatatt gg
//
