LOCUS       sequence-314388-        6229 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 6229)
  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 6229)
  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_mCherry_Hyg
FEATURES             Location/Qualifiers
     source          1..6229
                     /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..1784
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1219..1237)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1487..1506
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2205..2229
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(2320..2908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2400..2419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3079..3894)
                     /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(3079..3894)
                     /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     3802..3821
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3886..3905
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4185..4620
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4747..4769
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    4777..4953
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(5185..6210)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
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 gggatggtga
     1081 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1141 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1201 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1261 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1321 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1381 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1441 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1501 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1561 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1621 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1681 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1741 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaggagctc atatgaagat
     1801 gaagatgaaa tatttggtgt gtcaaataaa tagcttgtgt gcttaagttt gtgttttttt
     1861 cttggcttgt tgtgttatga atttgtggct ttttctaata ttaaatgaat gtaagatctc
     1921 attataatga ataaacaaat gtttctataa tccattgtga atgttttgtt ggatctcttc
     1981 tgcagcatat aactactgta tgtgctatgg tatggactat ggaatatgat taaagataag
     2041 gaattcagat cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa
     2101 acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctccgctcat
     2161 gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag gatatattgg
     2221 cgggtaaact aagtcgctgt atgtgtttgt ttgagatctc atgtgagcaa aaggccagca
     2281 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2341 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2401 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2461 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2521 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2581 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2641 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2701 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2761 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaagaa gagttggtag
     2821 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2881 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2941 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3001 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatgt
     3061 gtaacattgg tctagtgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3121 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3181 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3241 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3301 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     3361 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3421 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     3481 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3541 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttccctg ggatcgcagt
     3601 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     3661 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3721 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc ggtagattgt
     3781 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3841 gttggaattt aatcgcggcc ttgagcaaga cgtttcccgt tgaatatggc tcataacacc
     3901 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3961 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     4021 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     4081 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     4141 ctccctcact ttctggctgg atgatggggc gattcaggcg atccccatcc aacagcccgc
     4201 cgtcgagcgg gcttttttat ccccggaagc ctgtggatag agggtagtta tccacgtgaa
     4261 accgctaatg ccccgcaaag ccttgattca cggggctttc cggcccgctc caaaaactat
     4321 ccacgtgaaa tcgctaatca gggtacgtga aatcgctaat cggagtacgt gaaatcgcta
     4381 ataaggtcac gtgaaatcgc taatcaaaaa ggcacgtgag aacgctaata gccctttcag
     4441 atcaacagct tgcaaacacc cctcgctccg gcaagtagtt acagcaagta gtatgttcaa
     4501 ttagcttttc aattatgaat atatatatca attattggtc gcccttggct tgtggacaat
     4561 gcgctacgcg caccggctcc gcccgtggac aaccgcaagc ggttgcccac cgtcgagcgc
     4621 ctttgcccac aacccggcgg ccggccgcaa cagatcgttt tataaatttt tttttttgaa
     4681 aaagaaaaag cccgaaaggc ggcaacctct cgggcttctg gatttccgat ccccggaatt
     4741 agatcttggc aggatatatt gtggtgtaac gttatcgatc tggattttag tactggattt
     4801 tggttttagg aattagaaat tttattgata gaagtatttt acaaatacaa atacatacta
     4861 agggtttctt atatgctcaa cacatgagcg aaaccctata agaaccctaa tttcccttat
     4921 cgggaaacta ctcacacatt atttatggag aaaatagaga gagatagatt tgtagagaga
     4981 gactggtgat ttcagcgtac cgaattaatt cctgaatcac tgcgaccggc cctcccgcga
     5041 cccagccgag cgagcttagc gaactgtgga cgagaactgt gccaccaagc gtaaggccgt
     5101 tctctcgcat ttgccttgct aggctcgcgc gagttgctgg ctgaggcgtt ctcgaaatca
     5161 gctcttgttc ggtcggcatc tactctattc ctttgccctc ggacgagtgc tggggcgtcg
     5221 gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg cgcttctgcg
     5281 ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt cgcatcgacc
     5341 ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga gttggtcaag
     5401 accaatgcgg agcatatacg cccggagcct tggcgatcct gcaagctccg gatgcctccg
     5461 ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga agaagatgtt
     5521 ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga ccgctgttat
     5581 gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga ggtgccggac
     5641 ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga cggacgcact
     5701 gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa tcgcgcatat
     5761 gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc cgaacccgct
     5821 cgtctggcta agatcggccg cagcgatagc atccatagcc tccgcgaccg gctgaagaac
     5881 agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat
     5941 gcaataggtc aggctctcgc tgaactcccc aatgtcaagc acttccggaa tcgggagcgc
     6001 ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg cgcagctatt
     6061 tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag attcttcgcc
     6121 ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa acttctcgac
     6181 agacgtcgcg gtgagttcag gctttttcat aggggggatc agcttgggc
//
