LOCUS       sequence-460942-        8016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For transiently expressing fusion proteins with N terminal StrepIII 
            tags in plants (includes p19 silencing suppressor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8016)
  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 8016)
  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 
            Biotechnol J. 2025 Jun 26. doi: 10.1111/pbi.70233."
COMMENT     Sequence Label: pAVEC2_N-StrepIII
FEATURES             Location/Qualifiers
     source          1..8016
                     /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)"
     terminator      complement(3542..3790)
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     primer_bind     6152..6172
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    6221..6276
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             6295..6321
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6331..6357
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6367..6393
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(6424..6429,6430..6453,6454..6489,6490..6513)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(7050..7069)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7061..7870
                     /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(7128..7147)
                     /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 tctctagaaa
     3541 gcttatcttt aatcatattc catagtccat accatagcac atacagtagt tatatgctgc
     3601 agaagagatc caacaaaaca ttcacaatgg attatagaaa catttgttta ttcattataa
     3661 tgagatctta cattcattta atattagaaa aagccacaaa ttcataacac aacaagccaa
     3721 gaagaaaaca caaacttaag cacacaagct ttttatttga cacaccaaat atttcatctt
     3781 catcttcata tttactcgct ttctttttcg aaggtctcag taccttcagg gcatcctctt
     3841 gatacattac tttccacttc gattggggca agctgtagca gttcttgctt agaccgaatt
     3901 gccatctcac agagatgctg aagagttcgc gaccctccag aaacggtgat actaactcct
     3961 cgaaaccgaa tactataggt acatccgatc tggtcgaaac cgaaaaatcg agatgctgca
     4021 tagttaaccg aatctcccgt ccaagatcca aggactctgt gcagtgaagc ttccgtcctg
     4081 tcgtatctga gatatctctt aaatacaact ttcccgaaac cccagctttc cttgaaacca
     4141 aggggattat cttgattcga attagtctca tcgttatgta gccgccactc agtccaactc
     4201 ggactttcgt caggaagttt gaagggagaa gtggtacctc ctgatcctcc atcccaacgt
     4261 tcactgttag cttgttccct agcgtcgttt ccttgtatag ctcgttccat ctgttaatca
     4321 gaaaaactca gattaatcga caaattcgat cgcacaaact agaaactaac accagatcta
     4381 gatagaaatc acaaatcgaa gagtaattat tcgacaaaac tcaaattatt tgaacaaatc
     4441 ggatgatatc tatgaaaccc taatcgagaa ttaagatgat atctaacgat caaacccaga
     4501 aaatcgtctt cgatctaaga ttaacagaat ctaaaccaaa gaacatatac gaaattggga
     4561 tcgaacgaaa acaaaatcga agattttgag agaataagga acacagaaat ttaccttcat
     4621 cacggtagag agaattgaga gaaagttttt aagattttga gaaattgaaa tctgaattgt
     4681 gaagaagaag agctctttgg gtattgtttt atagaagaag aagaagaaaa gacgaggacg
     4741 actaggtcac gagaaagcta aggcggtgaa gcaatagcta ataataaaat gacacgtgta
     4801 ttgagcgttg tttacacgca aagttgtttt tggctaattg ccttattttt aggttgagga
     4861 aaagtatttg tgctttgagt tgataaacac gactcgtgtg tgccggctgc aaccactttg
     4921 acgccgttta ttactgactc gtcgactcga gactagagcc aagctgatct cctttgcccc
     4981 ggagatcacc atggacgact ttctctatct ctacgatcta ggaagaaagt tcgacggaga
     5041 aggtgacgat accatgttca ccaccgataa tgagaagatt agcctcttca atttcagaaa
     5101 gaatgctgac ccacagatgg ttagagaggc ttacgcggca ggtctcatca agacgatcta
     5161 cccgagtaat aatctccagg agatcaaata ccttcccaag aaggttaaag atgcagtcaa
     5221 aagattcagg actaactgca tcaagaacac agagaaagat atatttctca agatcagaag
     5281 tactattcca gtatggacga ttcaaggctt gcttcataaa ccaaggcaag taatagagat
     5341 tggagtctct aagaaagtag ttcctactga atcaaaggcc atggagtcaa aaattcagat
     5401 cgaggatcta acagaactcg ccgtgaagac tggcgaacag ttcatacaga gtcttttacg
     5461 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacactc tcgtctactc
     5521 caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag
     5581 ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag
     5641 gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat
     5701 cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
     5761 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
     5821 cactgacgta agggatgacg cacaatccca ctatccttcg caatgagact tttcaacaaa
     5881 gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa
     5941 ggacagtaga aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta
     6001 tcgttcaaga tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca
     6061 tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct
     6121 ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat
     6181 aaggaagttc atttcatttg gagagaggcc tatttttaca acaattacca acaacaacaa
     6241 acaacaaaca acattacaat tactatttac aattacaggc ctcccgggac catgtaccca
     6301 tacgatgtcc cagattatgc cgccggagcc tatccttatg atgttcctga ttacgctgct
     6361 ggtgcttacc cttacgatgt gcctgactac gccggtggtg gtagtggtgg tggaagtgga
     6421 ggatctgctt ggagccatcc tcagttcgag aaaggaggag gatctggagg aggttctggt
     6481 ggttctgctt ggtctcatcc acagtttgag aagtcaggaa gcgaaacacc aggaacatca
     6541 gaaagtgcaa caccagaatc tggatccggc tctgaaactc ctggtactag tgaatcggct
     6601 acccctgaat cagggtcctg atcacggcca tgctagagtc cgcaaaaatc accagtctct
     6661 ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag ttcccagata
     6721 agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt
     6781 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     6841 ccagtgacct ggtaacctgg caggatatat gcggttgtaa ttttggcagg atatatgcgg
     6901 ttgtaattca tttttattgt ctaaatttct gtatttgttt gtttgcaatt agatccacgt
     6961 tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     7021 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     7081 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     7141 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     7201 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     7261 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     7321 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     7381 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     7441 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     7501 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     7561 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     7621 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     7681 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     7741 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     7801 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     7861 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     7921 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     7981 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
