LOCUS       sequence-483211-        7057 bp    DNA     circular SYN 14-AUG-2025
DEFINITION  For transiently expressing fusion proteins with N terminal TurboID 
            for proximity labeling in plants (includes p19 silencing 
            suppressor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  AUTHORS   Greenhalgh R, Horner W, Klimpel KA, Chatterjee S, Scarpin MR, 
            Brunkard JO
  TITLE     Seamless, modular binary vectors for plant cell and molecular 
            biology.
  JOURNAL   Plant Biotechnol J. 2025 Jun 26. doi: 10.1111/pbi.70233.
  PUBMED    40574375
REFERENCE   2  (bases 1 to 7057)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Biotechnol J. 2025 Jun 26. doi: 10.1111/pbi.70233."
COMMENT     Sequence Label: pAVEC2_N-TurboID
FEATURES             Location/Qualifiers
     source          1..7057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             827..1456
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1885..2958
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3024..3218
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     4452..4472
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    4521..4576
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     primer_bind     complement(6091..6110)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6102..6911
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6169..6188)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgactga agaaccgagc
      721 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
      781 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
      841 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
      901 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
      961 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     1021 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     1081 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     1141 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     1201 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     1261 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     1321 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     1381 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     1441 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     1501 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     1561 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     1621 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     1681 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     1741 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     1801 aacgggcggt tggccaggcg taagcggctg ggttgcctgc cggccctgca atggcactgg
     1861 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
     1921 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     1981 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     2041 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
     2101 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     2161 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     2221 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     2281 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     2341 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     2401 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     2461 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     2521 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     2581 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     2641 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     2701 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
     2761 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
     2821 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
     2881 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
     2941 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
     3001 aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
     3061 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
     3121 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
     3181 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
     3241 gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
     3301 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
     3361 ccagcgcgcg gacaagctag gattggcagg atatattcag ttgtaaatgc gagctgttgg
     3421 ctggctggtg gcaggatata ttcagttgta aataaattga cgcttagaca acttaataac
     3481 acattgcgga cgtttttaat gtactgaatt aacgccgaat taattcgagc tctctagaat
     3541 cgtcgactcg agactagagc caagctgatc tcctttgccc cggagatcac catggacgac
     3601 tttctctatc tctacgatct aggaagaaag ttcgacggag aaggtgacga taccatgttc
     3661 accaccgata atgagaagat tagcctcttc aatttcagaa agaatgctga cccacagatg
     3721 gttagagagg cttacgcggc aggtctcatc aagacgatct acccgagtaa taatctccag
     3781 gagatcaaat accttcccaa gaaggttaaa gatgcagtca aaagattcag gactaactgc
     3841 atcaagaaca cagagaaaga tatatttctc aagatcagaa gtactattcc agtatggacg
     3901 attcaaggct tgcttcataa accaaggcaa gtaatagaga ttggagtctc taagaaagta
     3961 gttcctactg aatcaaaggc catggagtca aaaattcaga tcgaggatct aacagaactc
     4021 gccgtgaaga ctggcgaaca gttcatacag agtcttttac gactcaatga caagaagaaa
     4081 atcttcgtca acatggtgga gcacgacact ctcgtctact ccaagaatat caaagataca
     4141 gtctcagaag accaaagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc
     4201 ctcggattcc attgcccagc tatctgtcac ttcatcaaaa ggacagtaga aaaggaaggt
     4261 ggcacctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctgcc
     4321 gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt
     4381 ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt aagggatgac
     4441 gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg
     4501 gagagaggcc tatttttaca acaattacca acaacaacaa acaacaaaca acattacaat
     4561 tactatttac aattacaggc ctcccgggac catgaaagat aacactgttc cattgaaact
     4621 catcgctctc ctcgctaacg gtgagttcca ctctggtgag cagctcggtg agactttagg
     4681 tatgtctcgt gctgctatca acaagcacat ccagacttta cgtgattggg gagttgatgt
     4741 tttcactgtt ccaggaaagg gttactcttt accagagcca atcccattat tgaacgctaa
     4801 gcagatcttg ggtcagcttg atggaggatc tgttgctgtt cttccagttg ttgattctac
     4861 taaccagtac cttctcgatc gtattggaga gctcaagtct ggtgatgctt gcattgctga
     4921 gtaccagcag gcaggtcgtg gttctcgtgg taggaagtgg ttctctccat tcggtgcaaa
     4981 cctctacctt tctatgttct ggaggcttaa gaggggtcca gctgctattg gtcttggtcc
     5041 agttattggt attgttatgg ctgaggctct caggaagctc ggtgctgata aggttagggt
     5101 gaagtggcca aacgatctct acctccagga taggaagttg gctggtattt tggtggagct
     5161 tgctggaatt actggagatg ctgctcagat tgtgatcgga gctggaatca acgtggctat
     5221 gaggagggtc gaggagtctg tcgtcaacca gggatggatc actcttcagg aggctggtat
     5281 caaccttgat aggaacacac ttgctgctac acttatcagg gagcttaggg ctgctcttga
     5341 gttgttcgag caagagggtt tggctccata cttgccaagg tgggagaagt tggataactt
     5401 catcaacagg ccagtcaagt tgatcatcgg tgataaggag attttcggta tttctagggg
     5461 tattgataag caaggtgctt tgttgttgga gcaagatggt gttatcaagc catggatggg
     5521 tggtgagatc tctttgaggt ctgctgagaa ggggtcagga agcgaaacac caggaacatc
     5581 agaaagtgca acaccagaat ctggatccgg ctctgaaact cctggtacta gtgaatcggc
     5641 tacccctgaa tcagggtcct gatcacggcc atgctagagt ccgcaaaaat caccagtctc
     5701 tctctacaaa tctatctctc tctatttttc tccagaataa tgtgtgagta gttcccagat
     5761 aagggaatta gggttcttat agggtttcgc tcatgtgttg agcatataag aaacccttag
     5821 tatgtatttg tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa
     5881 tccagtgacc tggtaacctg gcaggatata tgcggttgta attttggcag gatatatgcg
     5941 gttgtaattc atttttattg tctaaatttc tgtatttgtt tgtttgcaat tagatccacg
     6001 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     6061 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     6121 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     6181 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     6241 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     6301 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     6361 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     6421 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     6481 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     6541 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     6601 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     6661 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     6721 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     6781 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     6841 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     6901 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     6961 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     7021 gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
