LOCUS       sequence-314390-        5476 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 5476)
  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 5476)
  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_Hyg
FEATURES             Location/Qualifiers
     source          1..5476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    962..986
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1077..1665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1157..1176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1836..2651)
                     /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(1836..2651)
                     /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     2559..2578
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2643..2662
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2942..3377
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    3504..3526
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    3534..3710
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(3942..4967)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(4987..5332)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(5024..5044)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 aacgctcaat acacgtgtca ttttattatt agctattgct tcaccgcctt agctttctcg
       61 tgacctagtc gtcctcgtct tttcttcttc ttcttctata aaacaatacc caaagacctc
      121 ttcttcttca caattcagat ttcaatttct caaaatctta aaaactttct ctcaattctc
      181 tctaccgtga tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt cgattttgtt
      241 ttcgttcgat cccaatttcg tatatgttct ttggtttaga ttctgttaat cttagatcga
      301 agacgatttt ctgggtttga tcgttagata tcatcttaat tctcgattag ggtttcatag
      361 atatcatccg atttgttcaa ataatttgag ttttgtcgaa taattactct tcgatttgtg
      421 atttctatct agatctggtg ttagtttcta gtttgtgcga tcgaatttgt cgattaatct
      481 gagtttttct gattaacaga ctagtggatc cgtcgacctc gagggtaccg ctcccgggta
      541 ggagctcata tgaagatgaa gatgaaatat ttggtgtgtc aaataaatag cttgtgtgct
      601 taagtttgtg tttttttctt ggcttgttgt gttatgaatt tgtggctttt tctaatatta
      661 aatgaatgta agatctcatt ataatgaata aacaaatgtt tctataatcc attgtgaatg
      721 ttttgttgga tctcttctgc agcatataac tactgtatgt gctatggtat ggactatgga
      781 atatgattaa agataaggaa ttcagatcgg ctgagtggct ccttcaacgt tgcggttctg
      841 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
      901 cttaattctc cgctcatgat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt
      961 ttgacaggat atattggcgg gtaaactaag tcgctgtatg tgtttgtttg agatctcatg
     1021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1561 ggaagaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1801 atctaaagta tatatgtgta acattggtct agtgattaga aaaactcatc gagcatcaaa
     1861 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1921 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1981 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2041 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2101 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2161 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2221 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2281 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2341 ttccctggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2401 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2461 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2521 tacaatcggt agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2581 tataaatcag catccatgtt ggaatttaat cgcggccttg agcaagacgt ttcccgttga
     2641 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2701 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     2761 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     2821 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     2881 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcgatc
     2941 cccatccaac agcccgccgt cgagcgggct tttttatccc cggaagcctg tggatagagg
     3001 gtagttatcc acgtgaaacc gctaatgccc cgcaaagcct tgattcacgg ggctttccgg
     3061 cccgctccaa aaactatcca cgtgaaatcg ctaatcaggg tacgtgaaat cgctaatcgg
     3121 agtacgtgaa atcgctaata aggtcacgtg aaatcgctaa tcaaaaaggc acgtgagaac
     3181 gctaatagcc ctttcagatc aacagcttgc aaacacccct cgctccggca agtagttaca
     3241 gcaagtagta tgttcaatta gcttttcaat tatgaatata tatatcaatt attggtcgcc
     3301 cttggcttgt ggacaatgcg ctacgcgcac cggctccgcc cgtggacaac cgcaagcggt
     3361 tgcccaccgt cgagcgcctt tgcccacaac ccggcggccg gccgcaacag atcgttttat
     3421 aaattttttt ttttgaaaaa gaaaaagccc gaaaggcggc aacctctcgg gcttctggat
     3481 ttccgatccc cggaattaga tcttggcagg atatattgtg gtgtaacgtt atcgatctgg
     3541 attttagtac tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca
     3601 aatacaaata catactaagg gtttcttata tgctcaacac atgagcgaaa ccctataaga
     3661 accctaattt cccttatcgg gaaactactc acacattatt tatggagaaa atagagagag
     3721 atagatttgt agagagagac tggtgatttc agcgtaccga attaattcct gaatcactgc
     3781 gaccggccct cccgcgaccc agccgagcga gcttagcgaa ctgtggacga gaactgtgcc
     3841 accaagcgta aggccgttct ctcgcatttg ccttgctagg ctcgcgcgag ttgctggctg
     3901 aggcgttctc gaaatcagct cttgttcggt cggcatctac tctattcctt tgccctcgga
     3961 cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag
     4021 acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg
     4081 attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc
     4141 tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccttgg cgatcctgca
     4201 agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc
     4261 ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag
     4321 tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg
     4381 tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc
     4441 tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga
     4501 tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg
     4561 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatagcatc catagcctcc
     4621 gcgaccggct gaagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
     4681 tgtgcacggc gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact
     4741 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
     4801 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
     4861 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
     4921 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatagg ggggatcagc
     4981 ttgggctgtc ctctccaaat gaaatgaact tccttatata gaggaagggt cttgcgaagg
     5041 atagtgggat tgtgcgtcat cccttacgtc agtggagatg tcacatcaat ccacttgctt
     5101 tgaagtcgtg gttggaacga cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
     5161 tttgggacca ctgtcggcag aggcatcttg aatgatagcc tttcctttat cgcaatgatg
     5221 gcatttgtag gagccacctt ccttttctac tgtcctttcg atgaagtgac agatagctgg
     5281 gcaatggaat ccgaggaggt ttcccgaaat taccctttgt tgaaaagtct caataaagct
     5341 tcgacgagtc agtaataaac ggcgtcaaag tggttgcagc cggcacacac gagtcgtgtt
     5401 tatcaactca aagcacaaat acttttcctc aacctaaaaa taaggcaatt agccaaaaac
     5461 aactttgcgt gtaaac
//
