LOCUS       sequence-461050-        7823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For stably 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 7823)
  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 7823)
  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: pAVEC1_N-StrepIII
FEATURES             Location/Qualifiers
     source          1..7823
                     /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(3545..3797)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(3807..4358)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(4364..4744)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     primer_bind     5959..5979
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    6028..6083
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             6102..6128
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6138..6164
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6174..6200
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(6231..6236,6237..6260,6261..6296,6297..6320)
                     /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(6857..6876)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6868..7677
                     /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(6935..6954)
                     /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 tcttctagag
     3541 atctgataat ttatttgaaa attcataaga aaagcaaacg ttacatgaat tgatgaaaca
     3601 atacaaagac agataaagcc acgcacattt aggatattgg ccgagattac tgaatattga
     3661 gtaagatcac ggaatttctg acaggagcat gtcttcaatt cagcccaaat ggcagttgaa
     3721 atactcaaac cgccccatat gcaggagcgg atcattcatt gtttgtttgg ttgcctttgc
     3781 caacatggga gtccaagatt ctgcagtcaa atctcggtga cgggcaggac cggacggggc
     3841 ggtaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt cccgtgcttg
     3901 aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg catgcgcacg
     3961 ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg tgcctccagg
     4021 gacttcagca ggtgggtgta gagcgtggag cccagtcccg tccgctggtg gcggggggag
     4081 acgtacacgg tcgactcggc cgtccagtcg taggcgttgc gtgccttcca ggggccagcg
     4141 taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata gcgctcccgc
     4201 agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg gaagttgacc
     4261 gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg
     4321 gcacggcgga tgtcggccgg gcgtcgttct gggctcatcg attcgatttg gtgtatcgag
     4381 attggttatg aaattcagat gctagtgtaa tgtattggta atttgggaag atataatagg
     4441 aagcaaggct atttatccat ttctgaaaag gcgaaatggc gtcaccgcga gcgtcacgcg
     4501 cattccgttc ttgctgtaaa gcgttgtttg gtacactttt gactagcgag gcttggcgtg
     4561 tcagcgtatc tattcaaaag tcgttaatgg ctgcggatca agaaaaagtt ggaatagaaa
     4621 cagaataccc gcgaaattca ggcccggttg ccatgtccta cacgccgaaa taaacgacca
     4681 aattagtaga aaaataaaaa ctgactcgga tacttacgtc acgtcttgcg cactgatttg
     4741 aaaaatctca gctcgaggac tagagccaag ctgatctcct ttgccccgga gatcaccatg
     4801 gacgactttc tctatctcta cgatctagga agaaagttcg acggagaagg tgacgatacc
     4861 atgttcacca ccgataatga gaagattagc ctcttcaatt tcagaaagaa tgctgaccca
     4921 cagatggtta gagaggctta cgcggcaggt ctcatcaaga cgatctaccc gagtaataat
     4981 ctccaggaga tcaaatacct tcccaagaag gttaaagatg cagtcaaaag attcaggact
     5041 aactgcatca agaacacaga gaaagatata tttctcaaga tcagaagtac tattccagta
     5101 tggacgattc aaggcttgct tcataaacca aggcaagtaa tagagattgg agtctctaag
     5161 aaagtagttc ctactgaatc aaaggccatg gagtcaaaaa ttcagatcga ggatctaaca
     5221 gaactcgccg tgaagactgg cgaacagttc atacagagtc ttttacgact caatgacaag
     5281 aagaaaatct tcgtcaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa
     5341 gatacagtct cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga
     5401 aacctcctcg gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag
     5461 gaaggtggca cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc
     5521 tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa
     5581 gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg
     5641 gatgacgcac aatcccacta tccttcgcaa tgagactttt caacaaaggg taatatcggg
     5701 aaacctcctc ggattccatt gcccagctat ctgtcacttc atcaaaagga cagtagaaaa
     5761 ggaaggtggc acctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc
     5821 ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga
     5881 agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag
     5941 ggatgacgca caatcccact atccttcgca agacccttcc tctatataag gaagttcatt
     6001 tcatttggag agaggcctat ttttacaaca attaccaaca acaacaaaca acaaacaaca
     6061 ttacaattac tatttacaat tacaggcctc ccgggaccat gtacccatac gatgtcccag
     6121 attatgccgc cggagcctat ccttatgatg ttcctgatta cgctgctggt gcttaccctt
     6181 acgatgtgcc tgactacgcc ggtggtggta gtggtggtgg aagtggagga tctgcttgga
     6241 gccatcctca gttcgagaaa ggaggaggat ctggaggagg ttctggtggt tctgcttggt
     6301 ctcatccaca gtttgagaag tcaggaagcg aaacaccagg aacatcagaa agtgcaacac
     6361 cagaatctgg atccggctct gaaactcctg gtactagtga atcggctacc cctgaatcag
     6421 ggtcctgatc acggccatgc tagagtccgc aaaaatcacc agtctctctc tacaaatcta
     6481 tctctctcta tttttctcca gaataatgtg tgagtagttc ccagataagg gaattagggt
     6541 tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     6601 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtgacctggt
     6661 aacctggcag gatatatgcg gttgtaattt tggcaggata tatgcggttg taattcattt
     6721 ttattgtcta aatttctgta tttgtttgtt tgcaattaga tccacgttgt gtctcaaaat
     6781 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     6841 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     6901 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     6961 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     7021 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     7081 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     7141 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     7201 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     7261 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     7321 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     7381 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     7441 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     7501 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     7561 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     7621 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     7681 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     7741 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     7801 gtagaaaaga tcaaaggatc ttc
//
