LOCUS       sequence-314407-        5764 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 5764)
  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 5764)
  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_Venus_Blp
FEATURES             Location/Qualifiers
     source          1..5764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(728..730,731..733,734..1447)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(773..794)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1034..1053)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1381..1402
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1868..1892
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1983..2571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2063..2082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2742..3557)
                     /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(2742..3557)
                     /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     3465..3484
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3549..3568
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3848..4283
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    4410..4432
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     polyA_signal    4440..4616
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(4848..5399)
                     /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(5419..5764)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(5456..5476)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 ataaagcttc gacgagtcag taataaacgg cgtcaaagtg gttgcagccg gcacacacga
       61 gtcgtgttta tcaactcaaa gcacaaatac ttttcctcaa cctaaaaata aggcaattag
      121 ccaaaaacaa ctttgcgtgt aaacaacgct caatacacgt gtcattttat tattagctat
      181 tgcttcaccg ccttagcttt ctcgtgacct agtcgtcctc gtcttttctt cttcttcttc
      241 tataaaacaa tacccaaaga cctcttcttc ttcacaattc agatttcaat ttctcaaaat
      301 cttaaaaact ttctctcaat tctctctacc gtgatcaagg taaatttctg tgttccttat
      361 tctctcaaaa tcttcgattt tgttttcgtt cgatcccaat ttcgtatatg ttctttggtt
      421 tagattctgt taatcttaga tcgaagacga ttttctgggt ttgatcgtta gatatcatct
      481 taattctcga ttagggtttc atagatatca tccgatttgt tcaaataatt tgagttttgt
      541 cgaataatta ctcttcgatt tgtgatttct atctagatct ggtgttagtt tctagtttgt
      601 gcgatcgaat ttgtcgatta atctgagttt ttctgattaa cagactagag ttaaccgggc
      661 tcaggcctgg cgcgccacta gtggatccat cgatagtact gtcgacctcg agggtaccgc
      721 tcccgggatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      781 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      841 cacctacggc aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg
      901 gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca
      961 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1021 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1081 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1141 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
     1201 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     1261 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1321 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1381 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1441 caagtaagag ctcatatgaa gatgaagatg aaatatttgg tgtgtcaaat aaatagcttg
     1501 tgtgcttaag tttgtgtttt tttcttggct tgttgtgtta tgaatttgtg gctttttcta
     1561 atattaaatg aatgtaagat ctcattataa tgaataaaca aatgtttcta taatccattg
     1621 tgaatgtttt gttggatctc ttctgcagca tataactact gtatgtgcta tggtatggac
     1681 tatggaatat gattaaagat aaggaattca gatcggctga gtggctcctt caacgttgcg
     1741 gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt
     1801 gactccctta attctccgct catgatcaga ttgtcgtttc ccgccttcag tttaaactat
     1861 cagtgtttga caggatatat tggcgggtaa actaagtcgc tgtatgtgtt tgtttgagat
     1921 ctcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2161 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2401 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2461 accttcggaa gaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2701 aaatcaatct aaagtatata tgtgtaacat tggtctagtg attagaaaaa ctcatcgagc
     2761 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2821 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2881 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2941 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     3001 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3061 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     3121 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     3181 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3241 gctgttttcc ctgggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3301 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3361 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3421 ttcccataca atcggtagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3481 tacccatata aatcagcatc catgttggaa tttaatcgcg gccttgagca agacgtttcc
     3541 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3601 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg
     3661 tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac gcatcttccc
     3721 gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg gtccacctac
     3781 aacaaagctc tcatcaaccg tggctccctc actttctggc tggatgatgg ggcgattcag
     3841 gcgatcccca tccaacagcc cgccgtcgag cgggcttttt tatccccgga agcctgtgga
     3901 tagagggtag ttatccacgt gaaaccgcta atgccccgca aagccttgat tcacggggct
     3961 ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa tcagggtacg tgaaatcgct
     4021 aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat cgctaatcaa aaaggcacgt
     4081 gagaacgcta atagcccttt cagatcaaca gcttgcaaac acccctcgct ccggcaagta
     4141 gttacagcaa gtagtatgtt caattagctt ttcaattatg aatatatata tcaattattg
     4201 gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc tccgcccgtg gacaaccgca
     4261 agcggttgcc caccgtcgag cgcctttgcc cacaacccgg cggccggccg caacagatcg
     4321 ttttataaat tttttttttt gaaaaagaaa aagcccgaaa ggcggcaacc tctcgggctt
     4381 ctggatttcc gatccccgga attagatctt ggcaggatat attgtggtgt aacgttatcg
     4441 atctggattt tagtactgga ttttggtttt aggaattaga aattttattg atagaagtat
     4501 tttacaaata caaatacata ctaagggttt cttatatgct caacacatga gcgaaaccct
     4561 ataagaaccc taatttccct tatcgggaaa ctactcacac attatttatg gagaaaatag
     4621 agagagatag atttgtagag agagactggt gatttcagcg taccgaatta attcctgaat
     4681 cactgcgacc ggccctcccg cgacccagcc gagcgagctt agcgaactgt ggacgagaac
     4741 tgtgccacca agcgtaaggc cgttctctcg catttgcctt gctaggctcg cgcgagttgc
     4801 tggctgaggc gttctcgaaa tcagctcttg ttcggtcggc atctacttca gatttcggtg
     4861 acgggcagga ccggacgggg cggcaccggc aggctgaagt ccagctgcca gaaacccacg
     4921 tcatgccagt tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg ggcatatccg
     4981 agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc gaccacgctc
     5041 ttgaagccct gtgcctccag ggacttcagc aggtgggtgt agagcgtgga gcccagtccc
     5101 gtccgctggt ggcgggggga gacgtacacg gtggactcgg ccgtccagtc gtaggcgttg
     5161 cgtgccttcc agggacccgc gtaggcgatg ccggcgacct cgccgtccac ctcggcgacg
     5221 agccagggat agcgctcccg cagacggacg aggtcgtccg tccactcctg cggttcctgc
     5281 ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat ggtgcagacc
     5341 gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc tgggctcata
     5401 ggggggatca gcttgggctg tcctctccaa atgaaatgaa cttccttata tagaggaagg
     5461 gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg tcagtggaga tgtcacatca
     5521 atccacttgc tttgaagtcg tggttggaac gacttctttt tccacgatgc tcctcgtggg
     5581 tgggggtcca tctttgggac cactgtcggc agaggcatct tgaatgatag cctttccttt
     5641 atcgcaatga tggcatttgt aggagccacc ttccttttct actgtccttt cgatgaagtg
     5701 acagatagct gggcaatgga atccgaggag gtttcccgaa attacccttt gttgaaaagt
     5761 ctca
//
