LOCUS       sequence-308940-        6697 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 6697)
  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 6697)
  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_LUC2_Blp
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    625..647
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    655..831
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1063..1614)
                     /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(1634..1979)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1671..1691)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    4780..4804
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4895..5483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4975..4994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5654..6469)
                     /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(5654..6469)
                     /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     6377..6396
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6461..6480
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
       61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
      121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
      181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
      241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
      301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
      361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
      421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
      481 ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta
      541 taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga
      601 tttccgatcc ccggaattag atcttggcag gatatattgt ggtgtaacgt tatcgatctg
      661 gattttagta ctggattttg gttttaggaa ttagaaattt tattgataga agtattttac
      721 aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa accctataag
      781 aaccctaatt tcccttatcg ggaaactact cacacattat ttatggagaa aatagagaga
      841 gatagatttg tagagagaga ctggtgattt cagcgtaccg aattaattcc tgaatcactg
      901 cgaccggccc tcccgcgacc cagccgagcg agcttagcga actgtggacg agaactgtgc
      961 caccaagcgt aaggccgttc tctcgcattt gccttgctag gctcgcgcga gttgctggct
     1021 gaggcgttct cgaaatcagc tcttgttcgg tcggcatcta cttcagattt cggtgacggg
     1081 caggaccgga cggggcggca ccggcaggct gaagtccagc tgccagaaac ccacgtcatg
     1141 ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc
     1201 ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa
     1261 gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg
     1321 ctggtggcgg ggggagacgt acacggtgga ctcggccgtc cagtcgtagg cgttgcgtgc
     1381 cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca
     1441 gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc
     1501 ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg
     1561 catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcataggggg
     1621 gatcagcttg ggctgtcctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt
     1681 gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt ggagatgtca catcaatcca
     1741 cttgctttga agtcgtggtt ggaacgactt ctttttccac gatgctcctc gtgggtgggg
     1801 gtccatcttt gggaccactg tcggcagagg catcttgaat gatagccttt cctttatcgc
     1861 aatgatggca tttgtaggag ccaccttcct tttctactgt cctttcgatg aagtgacaga
     1921 tagctgggca atggaatccg aggaggtttc ccgaaattac cctttgttga aaagtctcaa
     1981 taaagcttcg acgagtcagt aataaacggc gtcaaagtgg ttgcagccgg cacacacgag
     2041 tcgtgtttat caactcaaag cacaaatact tttcctcaac ctaaaaataa ggcaattagc
     2101 caaaaacaac tttgcgtgta aacaacgctc aatacacgtg tcattttatt attagctatt
     2161 gcttcaccgc cttagctttc tcgtgaccta gtcgtcctcg tcttttcttc ttcttcttct
     2221 ataaaacaat acccaaagac ctcttcttct tcacaattca gatttcaatt tctcaaaatc
     2281 ttaaaaactt tctctcaatt ctctctaccg tgatcaaggt aaatttctgt gttccttatt
     2341 ctctcaaaat cttcgatttt gttttcgttc gatcccaatt tcgtatatgt tctttggttt
     2401 agattctgtt aatcttagat cgaagtcgat tttctgggtt tgatcgttag atatcatctt
     2461 aattctcgat tagggtttca tagatatcat ccgatttgtt caaataattt gagttttgtc
     2521 gaataattac tcttcgattt gtgatttcta tctagatctg gtgttagttt ctagtttgtg
     2581 cgatcgaatt tgtcgattaa tctgagtttt tctgattaac agactagagt taaccgggct
     2641 caggcctggc gcgccactag tggatccatc gatagtactg tcgacctcga gggtaccgct
     2701 cccgggatgg aagatgctaa gaatattaag aagggaccag ctcctttcta cccactcgaa
     2761 gatggaacag caggtgaaca gttacacaag gctatgaaaa gatatgctct tgttcctggt
     2821 actatcgcat tcacagatgc tcatatcgag gtggatataa cttatgctga gtacttcgaa
     2881 atgtctgtta ggctcgctga agctatgaag agatacggat taaatacaaa ccacaggatt
     2941 gttgtgtgta gtgaaaattc tttgcaattt ttcatgcctg ttcttggagc tttgttcatc
     3001 ggtgtggctg ttgcaccagc taatgatata tataacgaga gagaacttct taattctatg
     3061 ggaatttctc aacctactgt tgtgttcgtg tctaagaaag gacttcaaaa gattttgaac
     3121 gttcagaaga aacttccaat tatccagaag attattatta tggattcaaa gacagattac
     3181 caaggtttcc agtcaatgta cacctttgtg actagtcatt tgcctcctgg attcaatgag
     3241 tacgatttcg ttccagaaag ttttgataga gataagacta ttgctctcat tatgaactct
     3301 tcaggatcta caggtcttcc taagggtgtg gcattgccac atagaaccgc ttgtgttagg
     3361 ttctctcacg caagagatcc tattttcgga aaccagataa ttccagatac agctatcttg
     3421 tcagttgtgc ctttccatca cggatttgga atgttcacta cactcggata ccttatttgt
     3481 ggttttaggg ttgtgctcat gtacagattc gaagaggaac tctttttaag atctttacaa
     3541 gattataaga tacagtcagc actcttagtt ccaacccttt tctctttctt tgctaagtca
     3601 actttgatcg ataagtacga tctttctaac ttgcatgaga ttgcaagtgg aggtgctcct
     3661 ctttctaaag aagtgggtga agcagttgct aagaggtttc acttgccagg aatcagacaa
     3721 ggatatggtc ttacagagac cacttctgct atcttgataa cccctgaagg agatgataaa
     3781 ccaggagcag ttggaaaggt tgttcctttc tttgaagcta aagttgtgga tctcgataca
     3841 ggaaagacct taggtgtgaa tcagagggga gagctttgcg ttagaggtcc tatgataatg
     3901 agtggatatg ttaataaccc agaagcaact aacgctctta tcgataaaga tggatggttg
     3961 cattcaggag atattgctta ctgggatgag gatgaacact ttttcatagt ggataggctc
     4021 aagtcattaa ttaagtataa gggataccaa gttgcacctg ctgagctcga aagtattctt
     4081 ttgcagcatc caaacatctt cgatgcagga gttgctggtc ttcctgatga tgatgctgga
     4141 gagctcccag ctgcagttgt ggttttagaa cacggaaaga ctatgacaga gaaagaaatt
     4201 gtggattacg ttgcatctca agttacaacc gctaagaaac ttagaggtgg tgtggttttt
     4261 gtggatgagg ttccaaaggg actcactggt aaattagatg ctagaaagat cagggaaatt
     4321 ttgatcaaag caaagaaagg aggaaagatt gctgtttgag agctcatatg aagatgaaga
     4381 tgaaatattt ggtgtgtcaa ataaatagct tgtgtgctta agtttgtgtt tttttcttgg
     4441 cttgttgtgt tatgaatttg tggctttttc taatattaaa tgaatgtaag atctcattat
     4501 aatgaataaa caaatgtttc tataatccat tgtgaatgtt ttgttggatc tcttctgcag
     4561 catataacta ctgtatgtgc tatggtatgg actatggaat atgattaaag ataaggaatt
     4621 cagatcggct gagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc
     4681 ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg ctcatgatca
     4741 gattgtcgtt tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt
     4801 aaactaagtc gctgtatgtg tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg
     4861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aagaagagtt ggtagctctt
     5401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgtgtaac
     5641 attggtctag tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     5701 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     5761 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     5821 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     5881 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     5941 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     6001 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     6061 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     6121 ctgaatcagg atattcttct aatacctgga atgctgtttt ccctgggatc gcagtggtga
     6181 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     6241 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     6301 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcggtag attgtcgcac
     6361 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     6421 aatttaatcg cggccttgag caagacgttt cccgttgaat atggctcata acaccccttg
     6481 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     6541 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     6601 ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc
     6661 aaaagttcaa aatcaccaac tggtccacct acaacaa
//
