LOCUS       sequence-460947-        6617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing fusion proteins with N terminal StrepIII tags in 
            plants .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6617)
  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 6617)
  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: pAVEC0_N-StrepIII
FEATURES             Location/Qualifiers
     source          1..6617
                     /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     4753..4773
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    4822..4877
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             4896..4922
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4932..4958
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4968..4994
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(5025..5030,5031..5054,5055..5090,5091..5114)
                     /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(5651..5670)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5662..6471
                     /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(5729..5748)
                     /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 gcaatgagac ttttcaacaa agggtaatat cgggaaacct
     4501 cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag aaaaggaagg
     4561 tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctgc
     4621 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
     4681 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
     4741 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
     4801 ggagagaggc ctatttttac aacaattacc aacaacaaca aacaacaaac aacattacaa
     4861 ttactattta caattacagg cctcccggga ccatgtaccc atacgatgtc ccagattatg
     4921 ccgccggagc ctatccttat gatgttcctg attacgctgc tggtgcttac ccttacgatg
     4981 tgcctgacta cgccggtggt ggtagtggtg gtggaagtgg aggatctgct tggagccatc
     5041 ctcagttcga gaaaggagga ggatctggag gaggttctgg tggttctgct tggtctcatc
     5101 cacagtttga gaagtcagga agcgaaacac caggaacatc agaaagtgca acaccagaat
     5161 ctggatccgg ctctgaaact cctggtacta gtgaatcggc tacccctgaa tcagggtcct
     5221 gatcacggcc atgctagagt ccgcaaaaat caccagtctc tctctacaaa tctatctctc
     5281 tctatttttc tccagaataa tgtgtgagta gttcccagat aagggaatta gggttcttat
     5341 agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa
     5401 atacttctat caataaaatt tctaattcct aaaaccaaaa tccagtgacc tggtaacctg
     5461 gcaggatata tgcggttgta attttggcag gatatatgcg gttgtaattc atttttattg
     5521 tctaaatttc tgtatttgtt tgtttgcaat tagatccacg ttgtgtctca aaatctctga
     5581 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     5641 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     5701 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     5761 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     5821 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     5881 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     5941 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     6001 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     6061 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     6121 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     6181 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     6241 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     6301 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     6361 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     6421 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     6481 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     6541 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     6601 aagatcaaag gatcttc
//
