LOCUS       sequence-482840-        7841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for TEV-mVenus-tagged proteins with FAST-Red 
            transgenic seed selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7841)
  AUTHORS   Schwartz S, Engstler C, Muhlbauer S, Windenbach E, Wunder T, Kunz 
            HH, Brandt B
  TITLE     
  JOURNAL   Plant J. 2025 Nov;124(3):e70544. doi: 10.1111/tpj.70544.
  PUBMED    41222448
REFERENCE   2  (bases 1 to 7841)
  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: "Plant J."; 
            date: "2025-11"; volume: "124(3):e70544. doi"; pages: " 
            10.1111/tpj.70544"
COMMENT     Sequence Label: pG20_TEV_mVenus_FR
FEATURES             Location/Qualifiers
     source          1..7841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..498
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    625..647
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     CDS             complement(1097..1792)
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     protein_bind    3770..3791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3806..3811,3812..3829,3830..3836)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3844..3860
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             3878..3889
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3896..4573
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      4618..4689
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4705..4732
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             4760..4780
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4784..4786,4787..4789,4790..5503)
                     /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(4829..4850)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5090..5109)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5437..5458
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5924..5948
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6039..6627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6119..6138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6798..7613)
                     /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(6798..7613)
                     /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     7521..7540
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7605..7624
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcgat
       61 ccccatccaa cagcccgccg tcgagcgggc ttttttatcc ccggaagcct gtggatagag
      121 ggtagttatc cacgtgaaac cgctaatgcc ccgcaaagcc ttgattcacg gggctttccg
      181 gcccgctcca aaaactatcc acgtgaaatc gctaatcagg gtacgtgaaa tcgctaatcg
      241 gagtacgtga aatcgctaat aaggtcacgt gaaatcgcta atcaaaaagg cacgtgagaa
      301 cgctaatagc cctttcagat caacagcttg caaacacccc tcgctccggc aagtagttac
      361 agcaagtagt atgttcaatt agcttttcaa ttatgaatat atatatcaat tattggtcgc
      421 ccttggcttg tggacaatgc gctacgcgca ccggctccgc ccgtggacaa ccgcaagcgg
      481 ttgcccaccg tcgagcgcct ttgcccacaa cccggcggcc ggccgcaaca gatcgtttta
      541 taaatttttt tttttgaaaa agaaaaagcc cgaaaggcgg caacctctcg ggcttctgga
      601 tttccgatcc ccggaattag atcttggcag gatatattgt ggtgtaacgt tatcactaaa
      661 tggagcaacc tactgttttt gaagtttggg ttgccatgta atcacttgca ataaatatat
      721 gtactaatga ctgcatcggc ttgcgtgtct aaatttgtag cttattgaaa actttttgtg
      781 gttaaagacg aatccagtta actaacataa gtttaacaaa acaagacctt ttccatatat
      841 agaccaccgt taccaactta caaagaagcc aaataaagta caacacaaca cttgcaagtt
      901 acaaaaagag aggaaaaaaa acatggtacc atttgacaac atatagaaac acacataaac
      961 attacttata acaaaaagat tcaaactcgg cttttattat ctccccctga tcaaattcta
     1021 aacactttat tcctcataaa ctaactcccc gacattggag ttattggact atgacgatga
     1081 catcagtggg gtaagctcac ttgtgcccca gtttgctagg gaggtcgcag tatctggcca
     1141 cagccacctc gtgctgctcg acgtaggttt ctttgtcggc ctccttgatt ctttccagtc
     1201 tgtggtccac atagtagacg ccgggcatct tgaggttctt agcgggtttc ttggatctgt
     1261 atgtggtctt gaagttgcag atcaggtggc ccccgcccac gagcttcagg gccatgtcgc
     1321 ttctgccttc caggccgccg tcagcggggt acagcatctc ggtgttggcc tcccagccga
     1381 gtgttttctt ctgcatcaca gggccgttgg atgggaagtt cacccctctg atcttgacgt
     1441 tgtagatgag gcagccgtcc tggaggctgg tgtcctgggt agcggtcagc acgcccccat
     1501 cttcgtatgt ggtgactctc tcccatgtga agccctcagg gaaggactgc ttaaagaagt
     1561 cggggatgcc ctgggtgtgg ttgatgaagg ttctgctgcc gtacatgaag ctggtagcca
     1621 ggatgtcgaa ggcgaagggg agagggccgc cctcgaccac cttgattctc atggtctggg
     1681 tgccctcgta gggcttgcct tcgccctcgg atgtgcactt gaagtggtgg ttgttcacgg
     1741 tgccctccat gtacagcttc atgtgcatgt tctccttaat cagctcgctc atagtagtgt
     1801 gctggccacc acgagtacgg tcacggtcat gttccccacc agtatgttgc tgtccgtagt
     1861 actgagctct gtctttcaga tcctgagctt tgcttcccaa cttcatcctt gcactgtcca
     1921 acttgtctga tccctgtggg tgctctcccg ttgcgtacct attcacaatc aataggtatc
     1981 ataccaagca ttttacctat atagatgata tacacagaca cattcaagct agctaggtca
     2041 aatttcgtat ttagttaacc aaatgttccc tagaatttgc aatttcttat tgttacattt
     2101 gttattcgac caaaaaaatt gatccaaagc tactggctca acacatgcat gcacatattg
     2161 cacaaaatta tgaagtaaga tgataaatgt gtgcttactt gtaaatccaa gagaaaacgg
     2221 ttatagcggc aatgccaaac cctccagagg aaagaaaacc ggtgatgagg agtgcaactg
     2281 tgatgagagc cgggacaagg attgggctga agataacgag cagaggtgtt gcaacagtca
     2341 aagctatgac agttccaaca agggtaaggc tggagagaac aaggagggaa ccaccagctg
     2401 tgacagcagt tgcagcttta gcaatctgcc tagacttgga gtagtcagat cctcgtccgg
     2461 acatctggta ctggtcacga tctcggccca tcatcgggta ctgatctctg ccgatgatat
     2521 cgtgatgggt tcctctagct gtatccgcca tttttttgtt cttgtttact agagagaatg
     2581 agattttgaa tgagagattg atcaaaaaga tgtgtatata ttttggaaga ggtaggagtg
     2641 tttgttagat atattaagaa aggagacgag aagtcacgtg tcaatggaga catcatgcaa
     2701 ttgtaggcca cgtgttaatg gcgtcaccat gcagccgctg catgtgttaa cgagaggagt
     2761 ctattcaaag acacgtttgt tttttgccta gctcagaatg tggcgttggc atcgaaccga
     2821 aacataaaac gcaaaacgaa cctttgggct cgtcagtatg ggtttgggcc ttcaatagat
     2881 tccttctata cggcccacat atccactatt ggcaacaaaa ttagaaaaat ttaggattta
     2941 gttttaaaat ttgttccgcg acattataaa gcttcgacga gtcagtaata aacggcgtca
     3001 aagtggttgc agccggcaca cacgagtcgt gtttatcaac tcaaagcaca aatacttttc
     3061 ctcaacctaa aaataaggca attagccaaa aacaactttg cgtgtaaaca acgctcaata
     3121 cacgtgtcat tttattatta gctattgctt caccgcctta gctttctcgt gacctagtcg
     3181 tcctcgtctt ttcttcttct tcttctataa aacaataccc aaagacctct tcttcttcac
     3241 aattcagatt tcaatttctc aaaatcttaa aaactttctc tcaattctct ctaccgtgat
     3301 caaggtaaat ttctgtgttc cttattctct caaaatcttc gattttgttt tcgttcgatc
     3361 ccaatttcgt atatgttctt tggtttagat tctgttaatc ttagatcgaa gtcgattttc
     3421 tgggtttgat cgttagatat catcttaatt ctcgattagg gtttcataga tatcatccga
     3481 tttgttcaaa taatttgagt tttgtcgaat aattactctt cgatttgtga tttctatcta
     3541 gatctggtgt tagtttctag tttgtgcgat cgaatttgtc gattaatctg agtttttctg
     3601 attaacagac tagagttaac cgggctcagg cctggcgcgc cactagtgga tccgatggtg
     3661 tcttcgattc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3721 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3781 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     3841 gaattgtgag cggataacaa tttcacacat actagagaaa gaggagaaat actagatggc
     3901 ttcctccgag gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt
     3961 taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca
     4021 gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc
     4081 cccgcagttc cagtacggtt ccaaggctta cgttaaacac ccggctgaca tcccggacta
     4141 cctgaaactg tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaggacgg
     4201 tggtgttgtt accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt
     4261 taaactgcgt ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaacgatggg
     4321 ttgggaagca tccaccgaac gtatgtaccc ggaggacggt gctctgaaag gtgaaatcaa
     4381 aatgcgtctg aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat
     4441 ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac
     4501 ctcccacaac gaggactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc
     4561 caccggtgct taataacgct gatagtgcta gtgtagatcg ctactagagc caggcatcaa
     4621 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     4681 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttataga
     4741 agacactgga cccgggagtg agaacctcta cttccaaggc tccatggtga gcaagggcga
     4801 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4861 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     4921 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     4981 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     5041 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     5101 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     5161 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     5221 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     5281 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     5341 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     5401 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     5461 cgccgccggg atcactctcg gcatggacga gctgtacaag taagagctca tatgaagatg
     5521 aagatgaaat atttggtgtg tcaaataaat agcttgtgtg cttaagtttg tgtttttttc
     5581 ttggcttgtt gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatctca
     5641 ttataatgaa taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct
     5701 gcagcatata actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg
     5761 aattcagatc ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa
     5821 cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg
     5881 atcagattgt cgtttcccgc cttcagttta aactatcagt gtttgacagg atatattggc
     5941 gggtaaacta agtcgctgta tgtgtttgtt tgagatctca tgtgagcaaa aggccagcaa
     6001 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6061 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6421 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaagaag agttggtagc
     6541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6661 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6721 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgtg
     6781 taacattggt ctagtgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     6841 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     6901 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     6961 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     7021 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     7081 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     7141 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     7201 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     7261 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttccctgg gatcgcagtg
     7321 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     7381 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     7441 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg gtagattgtc
     7501 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     7561 ttggaattta atcgcggcct tgagcaagac gtttcccgtt gaatatggct cataacaccc
     7621 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     7681 tgtgcaatgt aacatcagag attttgagac acaacgtggc tttgttgaat aaatcgaact
     7741 tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg ttccgtggca
     7801 aagcaaaagt tcaaaatcac caactggtcc acctacaaca a
//
