LOCUS       sequence-330283-        5755 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 5755)
  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 5755)
  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_Blp
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     170..189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    360..384
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(805..1515)
                     /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(1083..1102)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1352..1370
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        2243..2588
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2531..2551
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2608..3159
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    3391..3567
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3575..3597
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      3724..4159
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     complement(4439..4458)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4450..5265
                     /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             4450..5265
                     /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     complement(4523..4542)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(5436..5755,1..269)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
       61 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      121 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      181 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      241 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      301 tggccttttg ctggcctttt gctcacatga gatctcaaac aaacacatac agcgacttag
      361 tttacccgcc aatatatcct gtcaaacact gatagtttaa actgaaggcg ggaaacgaca
      421 atctgatcat gagcggagaa ttaagggagt cacgttatga cccccgccga tgacgcggga
      481 caagccgttt tacgtttgga actgacagaa ccgcaacgtt gaaggagcca ctcagccgat
      541 ctgaattcct tatctttaat catattccat agtccatacc atagcacata cagtagttat
      601 atgctgcaga agagatccaa caaaacattc acaatggatt atagaaacat ttgtttattc
      661 attataatga gatcttacat tcatttaata ttagaaaaag ccacaaattc ataacacaac
      721 aagccaagaa aaaaacacaa acttaagcac acaagctatt tatttgacac accaaatatt
      781 tcatcttcat cttcatatga gctcctactt gtacagctcg tccatgccgc cggtggagtg
      841 gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
      901 gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta
      961 ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat
     1021 ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat
     1081 ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
     1141 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     1201 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc
     1261 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag
     1321 gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt
     1381 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     1441 gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc
     1501 gcccttgctc accatcccgg gagcggtacc ctcgaggtcg acagtactat cgatggatcc
     1561 actagtggcg cgccaggcct gagcccggtt aactctagtc tgttaatcag aaaaactcag
     1621 attaatcgac aaattcgatc gcacaaacta gaaactaaca ccagatctag atagaaatca
     1681 caaatcgaag agtaattatt cgacaaaact caaattattt gaacaaatcg gatgatatct
     1741 atgaaaccct aatcgagaat taagatgata tctaacgatc aaacccagaa aatcgtcttc
     1801 gatctaagat taacagaatc taaaccaaag aacatatacg aaattgggat cgaacgaaaa
     1861 caaaatcgaa gattttgaga gaataaggaa cacagaaatt taccttgatc acggtagaga
     1921 gaattgagag aaagttttta agattttgag aaattgaaat ctgaattgtg aagaagaaga
     1981 ggtctttggg tattgtttta tagaagaaga agaagaaaag acgaggacga ctaggtcacg
     2041 agaaagctaa ggcggtgaag caatagctaa taataaaatg acacgtgtat tgagcgttgt
     2101 ttacacgcaa agttgttttt ggctaattgc cttattttta ggttgaggaa aagtatttgt
     2161 gctttgagtt gataaacacg actcgtgtgt gccggctgca accactttga cgccgtttat
     2221 tactgactcg tcgaagcttt attgagactt ttcaacaaag ggtaatttcg ggaaacctcc
     2281 tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa aaggaaggtg
     2341 gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat gcctctgccg
     2401 acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa gaagtcgttc
     2461 caaccacgac ttcaaagcaa gtggattgat gtgacatctc cactgacgta agggatgacg
     2521 cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca tttcatttgg
     2581 agaggacagc ccaagctgat cccccctatg agcccagaac gacgcccggc cgacatccgc
     2641 cgtgccaccg aggcggacat gccggcggtc tgcaccatcg tcaaccacta catcgagaca
     2701 agcacggtca acttccgtac cgagccgcag gaaccgcagg agtggacgga cgacctcgtc
     2761 cgtctgcggg agcgctatcc ctggctcgtc gccgaggtgg acggcgaggt cgccggcatc
     2821 gcctacgcgg gtccctggaa ggcacgcaac gcctacgact ggacggccga gtccaccgtg
     2881 tacgtctccc cccgccacca gcggacggga ctgggctcca cgctctacac ccacctgctg
     2941 aagtccctgg aggcacaggg cttcaagagc gtggtcgctg tcatcgggct gcccaacgac
     3001 ccgagcgtgc gcatgcacga ggcgctcgga tatgcccccc gcggcatgct gcgggcggcc
     3061 ggcttcaagc acgggaactg gcatgacgtg ggtttctggc agctggactt cagcctgccg
     3121 gtgccgcccc gtccggtcct gcccgtcacc gaaatctgaa gtagatgccg accgaacaag
     3181 agctgatttc gagaacgcct cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag
     3241 agaacggcct tacgcttggt ggcacagttc tcgtccacag ttcgctaagc tcgctcggct
     3301 gggtcgcggg agggccggtc gcagtgattc aggaattaat tcggtacgct gaaatcacca
     3361 gtctctctct acaaatctat ctctctctat tttctccata aataatgtgt gagtagtttc
     3421 ccgataaggg aaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac
     3481 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
     3541 ccaaaatcca gtactaaaat ccagatcgat aacgttacac cacaatatat cctgccaaga
     3601 tctaattccg gggatcggaa atccagaagc ccgagaggtt gccgcctttc gggctttttc
     3661 tttttcaaaa aaaaaaattt ataaaacgat ctgttgcggc cggccgccgg gttgtgggca
     3721 aaggcgctcg acggtgggca accgcttgcg gttgtccacg ggcggagccg gtgcgcgtag
     3781 cgcattgtcc acaagccaag ggcgaccaat aattgatata tatattcata attgaaaagc
     3841 taattgaaca tactacttgc tgtaactact tgccggagcg aggggtgttt gcaagctgtt
     3901 gatctgaaag ggctattagc gttctcacgt gcctttttga ttagcgattt cacgtgacct
     3961 tattagcgat ttcacgtact ccgattagcg atttcacgta ccctgattag cgatttcacg
     4021 tggatagttt ttggagcggg ccggaaagcc ccgtgaatca aggctttgcg gggcattagc
     4081 ggtttcacgt ggataactac cctctatcca caggcttccg gggataaaaa agcccgctcg
     4141 acggcgggct gttggatggg gatcgcctga atcgccccat catccagcca gaaagtgagg
     4201 gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc
     4261 tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca
     4321 aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg ttacattgca
     4381 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     4441 aggggtgtta tgagccatat tcaacgggaa acgtcttgct caaggccgcg attaaattcc
     4501 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     4561 gcgacaatct accgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     4621 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     4681 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     4741 accactgcga tcccagggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     4801 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4861 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     4921 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4981 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     5041 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     5101 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     5161 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     5221 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcact agaccaatgt
     5281 tacacatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5341 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5401 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5461 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5521 gagctaccaa ctcttcttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5581 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5641 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5701 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaa
//
