LOCUS       sequence-418448-        7557 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transiently expressing TurboID fused with a peroxisomal targeting 
            signal type I (PTS1) in planta .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7557)
  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 7557)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-TurboID-PTS1
FEATURES             Location/Qualifiers
     source          1..7557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             445..1236
                     /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      1747..1941
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2007..3080)
                     /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(3509..4138)
                     /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    4402..4426
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4995..5340
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     5283..5303
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             5350..5376
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5380..5406
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5410..5436
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5443..6399
                     /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"
     terminator      6463..6715
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6776..6800
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6923..7511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7003..7022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatca
       61 tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc ggagggtagc
      121 atgttgattg taacgatgac agagcgttgc tgcctgtgat caattcgggc acgaacccag
      181 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
      241 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
      301 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
      361 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
      421 cgccctaaaa caaagttaaa catcatgggg gaagcggtga tcgccgaagt atcgactcaa
      481 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
      541 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
      601 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
      661 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
      721 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
      781 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
      841 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
      901 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
      961 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     1021 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     1081 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     1141 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     1201 gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc tagctagaaa ttcgttcaag
     1261 ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga tgcactaagc acataattgc
     1321 tcacagccaa actatcaggt caagtctgct tttattattt ttaagcgtgc ataataagcc
     1381 ctacacaaat tgggagatat atcatgcatg accaaaatcc cttaacgtga gttttcgttc
     1441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt ctgccatttt tggggtgagg
     1501 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga gctcggtgcc ttgatgtggg
     1561 cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt
     1621 aggccagcca tttttgagcg gccagcggcc gcgataggcc gacgcgaagc ggcggggcgt
     1681 agggagcgca gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt gcgctggcca
     1741 gacagttatg cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag
     1801 gcggaaaaat cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca
     1861 atgtacggct ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc
     1921 ccaatgtacg tgctatccac aggaaagaag ctttttcgac ctttttcccc tgctagggca
     1981 atttgcccta gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg
     2041 ttgcggtagc gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac
     2101 tccggcaggt catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct
     2161 ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct
     2221 gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa
     2281 tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa
     2341 tcagctagct cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag
     2401 cggctaatca aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc
     2461 gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct
     2521 ttctgctttc cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg
     2581 aacacgcggc cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga
     2641 tgggaaaccg ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct
     2701 gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg
     2761 tcacgcttcg acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc
     2821 acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta
     2881 atcgacgacg caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc
     2941 gcttgccacg attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc
     3001 gcggccttca acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg
     3061 gatggtttgc gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg
     3121 gccggcagac aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc
     3181 attccacggc gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta
     3241 aaattcatct actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca
     3301 gatagcagct cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga
     3361 tcctccgccg gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc
     3421 aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt
     3481 ttgctcattt tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca
     3541 gcgcctggac ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg
     3601 cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct
     3661 cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg
     3721 acacgacaaa ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct
     3781 ccaccaggtc ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga
     3841 agtcggctgc cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca
     3901 ctacgaagtc gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc
     3961 cgacaacggt tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg
     4021 gatcggaatc gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg
     4081 ttgcgatggt cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc
     4141 gttccccttg cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg
     4201 caagctgttt tactcaaata cacatcacct ttttagacgg cggcgctcgg tttcttcagc
     4261 ggccaagctg gccggccagg ccgccagctt ggcatcagac aaaccggcca ggatttcatg
     4321 cagccgcacg gctgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
     4381 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag
     4441 acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggattac ggaagacaat
     4501 acggcggaga ttagcctttt caatttcaga aagaatgcta acccacagat ggttagagag
     4561 gcttacgcag cacgactcat caagacgatc tacccgagca ataatctcca ggaaatcaaa
     4621 taccttccca agaaggttaa agatgcagtc aaaagattca ggactaactg catcaagaac
     4681 acagagaaag atatatttct caagatcaga agtactattc cagtatggac gattcaaggc
     4741 ttgcttcaca aaccaaggca agtaatagag attggagtct ctaaaaaggt agttcccact
     4801 gaatcaaagg ccatggagtc aaagattcaa atagaggacc taacagaact cgccgtaaag
     4861 actggcgaac agttcataca gagtctctta cgactcaatg acaagaagaa aatcttcgtc
     4921 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
     4981 gagcaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
     5041 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
     5101 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
     5161 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaacacgt tccaaccacg
     5221 acttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc
     5281 cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagagaaca
     5341 cgcaccatgt acccgtatga tgttccggat tacgctggct atccctacga cgtgcccgac
     5401 tatgccgggt acccctatga cgtcccagac tacgcagcta gcaaagacaa tactgtgcct
     5461 ctgaagctga tcgctctcct ggctaatggc gagttccata gtggcgaaca gctgggagaa
     5521 accctgggca tgtccagggc cgctatcaac aagcacattc agactctgcg cgactggggc
     5581 gtggacgtgt tcaccgtgcc cggaaagggc tactctctgc ccgagcctat cccgctgctg
     5641 aacgctaaac agattctggg acagctggac ggcgggagcg tggcagtcct gcctgtggta
     5701 gactccacca atcagtacct gctggatcga atcggcgagc tgaagagtgg ggatgcttgc
     5761 attgcagaat atcagcaggc agggagagga agcagaggga ggaaatggtt ctctcctttt
     5821 ggagctaacc tgtacctgag tatgttttgg cgcctgaagc ggggaccagc agcaatcggc
     5881 ctgggcccgg tcatcggaat tgtcatggca gaagcgctgc gaaagctggg agcagacaag
     5941 gtgcgagtca aatggcccaa tgacctgtat ctgcaggata gaaagctggc aggcatcctg
     6001 gtggagctgg ccggaataac aggcgatgct gcacagatcg tcattggcgc cgggattaac
     6061 gtggctatga ggcgcgtgga ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa
     6121 gcagggatta acctggacag gaatactctg gccgctacgc tgatccgaga gctgcgggca
     6181 gccctggaac tgttcgagca ggaaggcctg gctccatatc tgccacggtg ggagaagctg
     6241 gataacttca tcaatagacc cgtgaagctg atcattgggg acaaagagat tttcgggatt
     6301 agccggggga ttgataaaca gggagccctg ctgctggaac aggacggagt tatcaaaccc
     6361 tggatgggcg gagaaatcag tctgcggtct gccgaaaagt ctaacttaca gcatcgtcgg
     6421 tctcatcttt agaagggcta agacctaagc tagcctagaa tcgatcgttc aaacatttgg
     6481 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt
     6541 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     6601 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     6661 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatctgagt
     6721 gacctaagct agcctagtgt cgggcttgtc ttctctggtt taaactatca gtgtttgaca
     6781 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     6841 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgccaaccac aggaaccgta
     6901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacg
//
