LOCUS       sequence-482854-        7844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for 3C-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 7844)
  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 7844)
  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_3C_mVenus_FR
FEATURES             Location/Qualifiers
     source          1..7844
                     /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..4783
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(4787..4789,4790..4792,4793..5506)
                     /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(4832..4853)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5093..5112)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5440..5461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5927..5951
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6042..6630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6122..6141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6801..7616)
                     /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(6801..7616)
                     /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     7524..7543
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7608..7627
                     /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 cccgggagtc tggaagttct gttccagggg ccctccatgg tgagcaaggg
     4801 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4861 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4921 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4981 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     5041 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5101 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     5161 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5221 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     5281 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     5341 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     5401 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     5461 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagagc tcatatgaag
     5521 atgaagatga aatatttggt gtgtcaaata aatagcttgt gtgcttaagt ttgtgttttt
     5581 ttcttggctt gttgtgttat gaatttgtgg ctttttctaa tattaaatga atgtaagatc
     5641 tcattataat gaataaacaa atgtttctat aatccattgt gaatgttttg ttggatctct
     5701 tctgcagcat ataactactg tatgtgctat ggtatggact atggaatatg attaaagata
     5761 aggaattcag atcggctgag tggctccttc aacgttgcgg ttctgtcagt tccaaacgta
     5821 aaacggcttg tcccgcgtca tcggcggggg tcataacgtg actcccttaa ttctccgctc
     5881 atgatcagat tgtcgtttcc cgccttcagt ttaaactatc agtgtttgac aggatatatt
     5941 ggcgggtaaa ctaagtcgct gtatgtgttt gtttgagatc tcatgtgagc aaaaggccag
     6001 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6061 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6481 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaag aagagttggt
     6541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6781 gtgtaacatt ggtctagtga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     6841 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     6901 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     6961 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     7021 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     7081 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     7141 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     7201 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     7261 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca
     7321 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     7381 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     7441 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcggtagatt
     7501 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     7561 atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg gctcataaca
     7621 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     7681 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     7741 acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg
     7801 gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaa
//
