LOCUS       sequence-314415-        7171 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 7171)
  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 7171)
  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_Hyg
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2660..2684
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2775..3363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2855..2874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3534..4349)
                     /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(3534..4349)
                     /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     4257..4276
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4341..4360
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4640..5075
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    5202..5224
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    5232..5408
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(5640..6665)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(6685..7030)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(6722..6742)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 aacaacgctc aatacacgtg tcattttatt attagctatt gcttcaccgc cttagctttc
       61 tcgtgaccta gtcgtcctcg tcttttcttc ttcttcttct ataaaacaat acccaaagac
      121 ctcttcttct tcacaattca gatttcaatt tctcaaaatc ttaaaaactt tctctcaatt
      181 ctctctaccg tgatcaaggt aaatttctgt gttccttatt ctctcaaaat cttcgatttt
      241 gttttcgttc gatcccaatt tcgtatatgt tctttggttt agattctgtt aatcttagat
      301 cgaagacgat tttctgggtt tgatcgttag atatcatctt aattctcgat tagggtttca
      361 tagatatcat ccgatttgtt caaataattt gagttttgtc gaataattac tcttcgattt
      421 gtgatttcta tctagatctg gtgttagttt ctagtttgtg cgatcgaatt tgtcgattaa
      481 tctgagtttt tctgattaac agactagagt taaccgggct caggcctggc gcgccactag
      541 tggatccatc gatagtactg tcgacctcga gggtaccgct cccgggatgg aagatgctaa
      601 gaatattaag aagggaccag ctcctttcta cccactcgaa gatggaacag caggtgaaca
      661 gttacacaag gctatgaaaa gatatgctct tgttcctggt actatcgcat tcacagatgc
      721 tcatatcgag gtggatataa cttatgctga gtacttcgaa atgtctgtta ggctcgctga
      781 agctatgaag agatacggat taaatacaaa ccacaggatt gttgtgtgta gtgaaaattc
      841 tttgcaattt ttcatgcctg ttcttggagc tttgttcatc ggtgtggctg ttgcaccagc
      901 taatgatata tataacgaga gagaacttct taattctatg ggaatttctc aacctactgt
      961 tgtgttcgtg tctaagaaag gacttcaaaa gattttgaac gttcagaaga aacttccaat
     1021 tatccagaag attattatta tggattcaaa gacagattac caaggtttcc agtcaatgta
     1081 cacctttgtg actagtcatt tgcctcctgg attcaatgag tacgatttcg ttccagaaag
     1141 ttttgataga gataagacta ttgctctcat tatgaactct tcaggatcta caggtcttcc
     1201 taagggtgtg gcattgccac atagaaccgc ttgtgttagg ttctctcacg caagagatcc
     1261 tattttcgga aaccagataa ttccagatac agctatcttg tcagttgtgc ctttccatca
     1321 cggatttgga atgttcacta cactcggata ccttatttgt ggttttaggg ttgtgctcat
     1381 gtacagattc gaagaggaac tctttttaag atctttacaa gattataaga tacagtcagc
     1441 actcttagtt ccaacccttt tctctttctt tgctaagtca actttgatcg ataagtacga
     1501 tctttctaac ttgcatgaga ttgcaagtgg aggtgctcct ctttctaaag aagtgggtga
     1561 agcagttgct aagaggtttc acttgccagg aatcagacaa ggatatggtc ttacagagac
     1621 cacttctgct atcttgataa cccctgaagg agatgataaa ccaggagcag ttggaaaggt
     1681 tgttcctttc tttgaagcta aagttgtgga tctcgataca ggaaagacct taggtgtgaa
     1741 tcagagggga gagctttgcg ttagaggtcc tatgataatg agtggatatg ttaataaccc
     1801 agaagcaact aacgctctta tcgataaaga tggatggttg cattcaggag atattgctta
     1861 ctgggatgag gatgaacact ttttcatagt ggataggctc aagtcattaa ttaagtataa
     1921 gggataccaa gttgcacctg ctgagctcga aagtattctt ttgcagcatc caaacatctt
     1981 cgatgcagga gttgctggtc ttcctgatga tgatgctgga gagctcccag ctgcagttgt
     2041 ggttttaaaa cacggaaaga ctatgacaga gaaagaaatt gtggattacg ttgcatctca
     2101 agttacaacc gctaagaaac ttagaggtgg tgtggttttt gtggatgagg ttccaaaggg
     2161 actcactggt aaattagatg ctagaaagat cagggaaatt ttgatcaaag caaagaaagg
     2221 aggaaagatt gctgtttgag agctcatatg aagatgaaga tgaaatattt ggtgtgtcaa
     2281 ataaatagct tgtgtgctta agtttgtgtt tttttcttgg cttgttgtgt tatgaatttg
     2341 tggctttttc taatattaaa tgaatgtaag atctcattat aatgaataaa caaatgtttc
     2401 tataatccat tgtgaatgtt ttgttggatc tcttctgcag catataacta ctgtatgtgc
     2461 tatggtatgg actatggaat atgattaaag ataaggaatt cagatcggct gagtggctcc
     2521 ttcaacgttg cggttctgtc agttccaaac gtaaaacggc ttgtcccgcg tcatcggcgg
     2581 gggtcataac gtgactccct taattctccg ctcatgatca gattgtcgtt tcccgccttc
     2641 agtttaaact atcagtgttt gacaggatat attggcgggt aaactaagtc gctgtatgtg
     2701 tttgtttgag atctcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2761 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2821 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2881 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2941 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3001 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3061 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3121 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3181 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3241 ctgaagccag ttaccttcgg aagaagagtt ggtagctctt gatccggcaa acaaaccacc
     3301 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3361 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3421 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3481 aaatgaagtt ttaaatcaat ctaaagtata tatgtgtaac attggtctag tgattagaaa
     3541 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     3601 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     3661 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     3721 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     3781 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     3841 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     3901 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     3961 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     4021 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     4081 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     4141 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     4201 ggcgcatcgg gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg
     4261 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag
     4321 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     4381 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     4441 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc
     4501 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac
     4561 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat
     4621 ggggcgattc aggcgatccc catccaacag cccgccgtcg agcgggcttt tttatccccg
     4681 gaagcctgtg gatagagggt agttatccac gtgaaaccgc taatgccccg caaagccttg
     4741 attcacgggg ctttccggcc cgctccaaaa actatccacg tgaaatcgct aatcagggta
     4801 cgtgaaatcg ctaatcggag tacgtgaaat cgctaataag gtcacgtgaa atcgctaatc
     4861 aaaaaggcac gtgagaacgc taatagccct ttcagatcaa cagcttgcaa acacccctcg
     4921 ctccggcaag tagttacagc aagtagtatg ttcaattagc ttttcaatta tgaatatata
     4981 tatcaattat tggtcgccct tggcttgtgg acaatgcgct acgcgcaccg gctccgcccg
     5041 tggacaaccg caagcggttg cccaccgtcg agcgcctttg cccacaaccc ggcggccggc
     5101 cgcaacagat cgttttataa attttttttt ttgaaaaaga aaaagcccga aaggcggcaa
     5161 cctctcgggc ttctggattt ccgatccccg gaattagatc ttggcaggat atattgtggt
     5221 gtaacgttat cgatctggat tttagtactg gattttggtt ttaggaatta gaaattttat
     5281 tgatagaagt attttacaaa tacaaataca tactaagggt ttcttatatg ctcaacacat
     5341 gagcgaaacc ctataagaac cctaatttcc cttatcggga aactactcac acattattta
     5401 tggagaaaat agagagagat agatttgtag agagagactg gtgatttcag cgtaccgaat
     5461 taattcctga atcactgcga ccggccctcc cgcgacccag ccgagcgagc ttagcgaact
     5521 gtggacgaga actgtgccac caagcgtaag gccgttctct cgcatttgcc ttgctaggct
     5581 cgcgcgagtt gctggctgag gcgttctcga aatcagctct tgttcggtcg gcatctactc
     5641 tattcctttg ccctcggacg agtgctgggg cgtcggtttc cactatcggc gagtacttct
     5701 acacagccat cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc
     5761 ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa
     5821 ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat atacgcccgg
     5881 agccttggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc
     5941 atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg ggaatccccg
     6001 aacatcgcct cgctccagtc aatgaccgct gttatgcggc cattgtccgt caggacattg
     6061 ttggagccga aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc
     6121 atcagctcat cgagagcctg cgcgacggac gcactgacgg tgtcgtccat cacagtttgc
     6181 cagtgataca catggggatc agcaatcgcg catatgaaat cacgccatgt agtgtattga
     6241 ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg
     6301 atagcatcca tagcctccgc gaccggctga agaacagcgg gcagttcggt ttcaggcagg
     6361 tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat aggtcaggct ctcgctgaac
     6421 tccccaatgt caagcacttc cggaatcggg agcgcggccg atgcaaagtg ccgataaaca
     6481 taacgatctt tgtagaaacc atcggcgcag ctatttaccc gcaggacata tccacgccct
     6541 cctacatcga agctgaaagc acgagattct tcgccctccg agagctgcat caggtcggag
     6601 acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt
     6661 ttcatagggg ggatcagctt gggctgtcct ctccaaatga aatgaacttc cttatataga
     6721 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatgtc
     6781 acatcaatcc acttgctttg aagtcgtggt tgtaacgact tctttttcca cgatgctcct
     6841 cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa tgatagcctt
     6901 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat
     6961 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta ccctttgttg
     7021 aaaagtctca ataaagcttc gacgagtcag taataaacgg cgtcaaagtg gttgcagccg
     7081 gcacacacga gtcgtgttta tcaactcaaa gcacaaatac ttttcctcaa cctaaaaata
     7141 aggcaattag ccaaaaacaa ctttgcgtgt a
//
