LOCUS       sequence-418443-        8497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transiently expressing and visualizing the subcellular localization 
            of TurboID fused with a Mitochondrial transit peptide (mTP) in 
            planta .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8497)
  AUTHORS   Tan H, Zhou Y, Dinius E, Lozano-Duran R
  TITLE     The Ti-TAN plasmid toolbox for TurboID-based proximity labeling 
            assays in Nicotiana benthamiana.
  JOURNAL   J Integr Plant Biol. 2024 Jan 31. doi: 10.1111/jipb.13610.
  PUBMED    38294100
REFERENCE   2  (bases 1 to 8497)
  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: "J Integr 
            Plant Biol. 2024 Jan 31. doi: 10.1111/jipb.13610."
COMMENT     Sequence Label: pGG-mTP-TurboID-GFP
FEATURES             Location/Qualifiers
     source          1..8497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             446..1237
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1748..1942
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2008..3081)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(3510..4139)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    4403..4427
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4996..5341
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5284..5304
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             complement(5386..5397)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5582..5608
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5612..5638
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5642..5668
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5675..6631
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             join(6683..6685,6686..7399)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(6986..7005)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7333..7354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      7404..7656
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    7717..7741
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(7864..8452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7944..7963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
       61 atcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag
      121 catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaattcggg cacgaaccca
      181 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
      241 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
      301 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
      361 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
      421 tcgccctaaa acaaagttaa acatcatggg ggaagcggtg atcgccgaag tatcgactca
      481 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
      541 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
      601 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
      661 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
      721 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
      781 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
      841 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
      901 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
      961 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     1021 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     1081 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     1141 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     1201 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctagctagaa attcgttcaa
     1261 gccgacgccg cttcgcggcg cggcttaact caagcgttag atgcactaag cacataattg
     1321 ctcacagcca aactatcagg tcaagtctgc ttttattatt tttaagcgtg cataataagc
     1381 cctacacaaa ttgggagata tatcatgcat gaccaaaatc ccttaacgtg agttttcgtt
     1441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tctgccattt ttggggtgag
     1501 gccgttcgcg gccgaggggc gcagcccctg gggggatggg agctcggtgc cttgatgtgg
     1561 gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg
     1621 taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg
     1681 tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc
     1741 agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta
     1801 ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc
     1861 aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt
     1921 cccaatgtac gtgctatcca caggaaagaa gctttttcga cctttttccc ctgctagggc
     1981 aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag
     2041 gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta
     2101 ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc
     2161 tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc
     2221 tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta
     2281 atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca
     2341 atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta
     2401 gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt
     2461 cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc
     2521 tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg
     2581 gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag
     2641 atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc
     2701 tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg
     2761 gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc
     2821 cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct
     2881 aatcgacgac gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc
     2941 cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg
     3001 cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc
     3061 ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag
     3121 ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg
     3181 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
     3241 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
     3301 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
     3361 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
     3421 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
     3481 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
     3541 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
     3601 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
     3661 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
     3721 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
     3781 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
     3841 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
     3901 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
     3961 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
     4021 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
     4081 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
     4141 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
     4201 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
     4261 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
     4321 gcagccgcac ggctgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
     4381 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
     4441 gacaacttaa taacacattg cggacgtttt taatgtactg gggtggatta cggaagacaa
     4501 tacggcggag attagccttt tcaatttcag aaagaatgct aacccacaga tggttagaga
     4561 ggcttacgca gcacgactca tcaagacgat ctacccgagc aataatctcc aggaaatcaa
     4621 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
     4681 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
     4741 cttgcttcac aaaccaaggc aagtaataga gattggagtc tctaaaaagg tagttcccac
     4801 tgaatcaaag gccatggagt caaagattca aatagaggac ctaacagaac tcgccgtaaa
     4861 gactggcgaa cagttcatac agagtctctt acgactcaat gacaagaaga aaatcttcgt
     4921 caacatggtg gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcaga
     4981 agagcaaagg gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt
     5041 ccattgccca gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta
     5101 caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg
     5161 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaacacg ttccaaccac
     5221 gacttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
     5281 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac
     5341 acgcaccatg gcaatggctg ttttccgtcg cgaagggagg cgtctcctcc cttcaatcgc
     5401 cgctcgccca atcgctgcta tccgatctcc tctctcttct gaccaggagg aaggacttct
     5461 tggagttcga tctatctcaa ctcaagtggt gcgtaaccgc atgaagagtg ttaagaacat
     5521 ccaaaagatc acaaaggcaa tgaagatggt tgctgcttcc aagcttagag cagttcaaat
     5581 gtacccgtat gatgttccgg attacgctgg ctatccctac gacgtgcccg actatgccgg
     5641 gtacccctat gacgtcccag actacgcagc tagcaaagac aatactgtgc ctctgaagct
     5701 gatcgctctc ctggctaatg gcgagttcca tagtggcgaa cagctgggag aaaccctggg
     5761 catgtccagg gccgctatca acaagcacat tcagactctg cgcgactggg gcgtggacgt
     5821 gttcaccgtg cccggaaagg gctactctct gcccgagcct atcccgctgc tgaacgctaa
     5881 acagattctg ggacagctgg acggcgggag cgtggcagtc ctgcctgtgg tagactccac
     5941 caatcagtac ctgctggatc gaatcggcga gctgaagagt ggggatgctt gcattgcaga
     6001 atatcagcag gcagggagag gaagcagagg gaggaaatgg ttctctcctt ttggagctaa
     6061 cctgtacctg agtatgtttt ggcgcctgaa gcggggacca gcagcaatcg gcctgggccc
     6121 ggtcatcgga attgtcatgg cagaagcgct gcgaaagctg ggagcagaca aggtgcgagt
     6181 caaatggccc aatgacctgt atctgcagga tagaaagctg gcaggcatcc tggtggagct
     6241 ggccggaata acaggcgatg ctgcacagat cgtcattggc gccgggatta acgtggctat
     6301 gaggcgcgtg gaggaaagcg tggtcaatca gggctggatc acactgcagg aagcagggat
     6361 taacctggac aggaatactc tggccgctac gctgatccga gagctgcggg cagccctgga
     6421 actgttcgag caggaaggcc tggctccata tctgccacgg tgggagaagc tggataactt
     6481 catcaataga cccgtgaagc tgatcattgg ggacaaagag attttcggga ttagccgggg
     6541 gattgataaa cagggagccc tgctgctgga acaggacgga gttatcaaac cctggatggg
     6601 cggagaaatc agtctgcggt ctgccgaaaa gaagggaggt ggaggaggtt ctggaggcgg
     6661 tggaagtggt ggcggaggta gcgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     6721 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     6781 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     6841 gcccgtgccc tggcccaccc tcgtgaccac cttcacctac ggcgtgcagt gcttcagccg
     6901 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     6961 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     7021 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     7081 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     7141 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     7201 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     7261 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     7321 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat
     7381 ggacgagctg tacaagtaaa atcgatcgtt caaacatttg gcaataaagt ttcttaagat
     7441 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     7501 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     7561 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     7621 aattatcgcg cgcggtgtca tctatgttac tagatctgag tgacctaagc tagcctagtg
     7681 tcgggcttgt cttctctggt ttaaactatc agtgtttgac aggatatatt ggcgggtaaa
     7741 cctaagagaa aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt
     7801 atccgttcgt ccatttgtat gtgccaacca caggaaccgt aaaaaggccg cgttgctggc
     7861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     8101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     8161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     8221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     8281 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     8341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     8401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     8461 tttgatcttt tctacggggt ctgacgctca gtggaac
//
