LOCUS       sequence-176885-        9319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescence complementation assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9319)
  AUTHORS   Frei dit Frey N, Mbengue M, Kwaaitaal M, Nitsch L, Altenbach D, 
            Haweker H, Lozano-Duran R, Njo MF, Beeckman T, Huettel B, Borst JW, 
            Panstruga R, Robatzek S
  TITLE     Plasma membrane calcium ATPases are important components of 
            receptor-mediated signaling in plant immune responses and 
            development.
  JOURNAL   Plant Physiol. 2012 Jun;159(2):798-809. doi: 10.1104/pp.111.192575. 
            Epub 2012 Apr 25.
  PUBMED    22535420
REFERENCE   2  (bases 1 to 9319)
  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"; doi: 
            "10.1104/pp.111.192575"; journalName: "Plant Physiol"; date: 
            "2012-06"; volume: "159"; issue: "2"; pages: "798-809"
COMMENT     Sequence Label: FLS2-YFPn
FEATURES             Location/Qualifiers
     source          1..9319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..345
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     289..309
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     347..363
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     348..364
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    380..404
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3960..3984)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3992..3994,3995..3997,3998..4456)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(4037..4058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4298..4317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4748..4772
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4884..5593
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5666..5757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5758..5826,5827..6618)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5976..5995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6789..7377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7278..7297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7689..7713
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(7807..8059)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8079..8630)
                     /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(8672..8851)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttc atcgaaagga cagtagaaaa ggaagatggc ttctacaaat gccatcattg
      121 cgataaagga aaggctatcg ttcaagatgc ctctaccgac agtggtccca aagatggacc
      181 cccacccacg aggaacatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
      241 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag aggacctcga ggtcgacggt
      361 atcgataagc ttcccatcaa caagtttgta caaaaaagca ggctccgcgg ccgccccctt
      421 caccatgaag ttactctcaa agaccttttt gatattaact ctcaccttct tcttctttgg
      481 cattgcacta gcgaaacaga gctttgaacc agagatcgaa gctttgaaat ccttcaagaa
      541 tggtatttcc aacgaccctt taggagtatt atcagattgg accatcatcg gttcgttacg
      601 acactgtaat tggaccggaa tcacctgcga tagtaccgga catgtagtct cggtttcctt
      661 gctggagaag caacttgaag gtgttctgtc tccagccata gcgaatctca cctatctcca
      721 ggttcttgat ctcacttcaa atagttttac cggcaaaata ccggctgaaa taggaaagtt
      781 aaccgagctt aaccagctta ttctgtacct aaactatttc tctggttcga ttccttctgg
      841 aatctgggag cttaagaata ttttctatct tgatcttaga aataatttgt tgtccggtga
      901 tgttcctgag gaaatctgca aaaccagttc tttggtattg attgggtttg attacaacaa
      961 cttaaccggg aaaataccag aatgcttagg agatttggtt catctccaaa tgtttgtagc
     1021 agctggtaac catttaactg gttcgattcc ggtatcaatt ggtactctgg ctaatttaac
     1081 ggatttagac ctgagtggta accagttaac cggaaaaata ccgagagatt ttggaaatct
     1141 cttgaactta cagtctctcg ttttaactga aaacttgttg gaaggagata taccagctga
     1201 gatcggaaac tgctcgagct tggtccaact tgagctttac gataaccagt taaccgggaa
     1261 aataccagct gaattaggga atttggttca gctgcaagca ctccggatat acaagaacaa
     1321 acttacttct tcaattccat cttcattgtt ccggttaact cagttaaccc atttggggtt
     1381 atcagaaaac catttggttg gaccgatatc agaagaaatc ggttttcttg agtcacttga
     1441 agtcctcaca cttcattcca acaacttcac aggagagttt ccacagtcca tcacaaactt
     1501 gaggaacttg acagtcctaa cggtggggtt caataatatt tccggtgagc tcccggcgga
     1561 tctagggctt cttacaaacc ttcggaacct ttcagcgcac gacaatcttc ttaccggacc
     1621 aataccttcc agcataagta actgcaccgg tcttaaactc ctggacctgt ctcacaacca
     1681 aatgactggc gagatcccgc ggggtttcgg aaggatgaat cttacgttca tttctattgg
     1741 gaggaatcat ttcaccggtg aaattccaga tgatatcttc aactgttcaa acttggaaac
     1801 tcttagtgtg gcagataaca acttaacagg aactctcaag ccattaattg ggaagcttca
     1861 aaaactcagg attttgcaag tttcatataa ctctctcact ggaccgattc ctcgagaaat
     1921 cgggaatctg aaagatttga atatcttgta ccttcactct aatggtttca cagggagaat
     1981 cccgagagag atgtcgaatc tcactctcct ccaggggcta aggatgtatt caaatgatct
     2041 tgaaggtcca attcctgaag aaatgtttga tatgaagcta ctctcagttc ttgatctttc
     2101 caacaacaaa ttctcaggtc aaattcctgc cttgttctcc aagcttgaat cgcttaccta
     2161 cttgagtctt caaggaaaca aattcaacgg gtctatccct gcaagcctta agtcgctttc
     2221 gcttctcaac acattcgata tctccgacaa tcttctcact ggaaccatcc ctggagagct
     2281 gttagcttct ttgaaaaaca tgcagcttta cctcaacttc tcaaacaact tgttgactgg
     2341 aaccatccca aaggagcttg gaaagcttga aatggttcaa gaaatcgacc tttcaaacaa
     2401 tctcttttct gggtctattc caagatcttt acaggcctgc aaaaatgtgt tcacactgga
     2461 tttttcgcag aacaatctct cgggtcatat accagatgaa gtcttccaag gcatggatat
     2521 gatcataagc ttgaaccttt caaggaacag tttctctgga gaaatccctc agagcttcgg
     2581 gaacatgacg catttggtct ccttggatct ctctagtaac aatctcactg gtgaaattcc
     2641 agagagtctc gccaatcttt cgactctgaa acatctcaaa ctagcttcaa acaacctcaa
     2701 aggccatgtt cctgaatccg gggtgttcaa aaacatcaac gcctctgatc taatgggaaa
     2761 cacagatctc tgtggtagca agaagcctct caagccatgt acgatcaagc agaagtcgag
     2821 ccacttctcg aagagaacca gagtcatcct gattattctt ggatcagccg cggctcttct
     2881 tcttgtcctg cttcttgttc tgattctaac ctgttgcaag aaaaaagaaa aaaagattga
     2941 aaattcatca gagtcctcat taccggattt ggattcagct ctgaaactga agagatttga
     3001 accaaaagag ttggagcaag caacagattc attcaacagt gccaacatca ttggctcaag
     3061 cagcttaagc acagtgtaca aaggtcagct agaagatggg acagtgattg cagtaaaagt
     3121 attgaatcta aaggaattct ctgcagaatc agacaagtgg ttctacacag aagctaaaac
     3181 attgagccaa ctaaaacatc gaaacctggt gaagatctta gggtttgcgt gggaaagcgg
     3241 caaaacgaaa gctttagtgc ttccatttat ggagaatgga aacttggagg acaccattca
     3301 cggctctgca gcaccgattg ggtcgctttt agaaaaaatc gatctttgtg ttcatatcgc
     3361 aagcggaatc gattatcttc attctggata tggttttccc atcgttcatt gtgatctgaa
     3421 gccagctaat atactccttg acagtgaccg cgttgctcac gtaagcgatt ttggaactgc
     3481 tcggattcta ggtttccgcg aagatggaag caccacagct tcaacatcag ccttcgaggg
     3541 tacaattgga tacttagctc cagagtttgc ttatatgagg aaagtgacaa caaaagccga
     3601 tgtattcagc ttcgggatca taatgatgga gctgatgacg aaacagagac caacttcgtt
     3661 gaatgatgaa gattcacaag acatgacttt gcgccaattg gtggagaaat cgattggaaa
     3721 tggaagaaaa gggatggtta gggttcttga tatggaactc ggggactcta ttgtttctct
     3781 gaaacaggaa gaggctattg aagactttct gaagctttgt ttgttctgta caagctctag
     3841 acctgaagat cgacctgata tgaacgagat tcttacacat ctgatgaaac ttagaggcaa
     3901 agcgaattca tttcgagaag atcgtaacga ggatcgagaa gttaagggtg ggcgcgccga
     3961 cccagctttc ttgtacaaag tggttgatgg gatggtgagc aagggcgagg agctgttcac
     4021 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4081 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac
     4141 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca
     4201 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4261 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4321 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4381 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4441 cgtctatatc atggcctaat ctagagtccg caaaaatcac cagtctctct ctacaaatct
     4501 atctctctct atttttctcc agaataatgt gtgagtagtt cccagataag ggaattaggg
     4561 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     4621 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgaccggg
     4681 cggccgccac cgcggtggag ggggatcaga ttgtcgtttc ccgccttcag tttaaactat
     4741 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     4801 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgtacatc
     4861 accgacgagc aaggcaagac cgagcgcctt tccgacgctc accgggctgg ttgccctcgc
     4921 cgctgggctg gcggccgtct atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg
     4981 tgcgagacac cggccgccgg cgttgtggat acctcgcgga aaacttggcc ctcactgaca
     5041 gatgaggggc ggacgttgac acttgagggg ccgactcacc cggcgcggcg ttgacagatg
     5101 aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa
     5161 acgcctgatt ttacgcgagt ttcccacaga tgatgtggac aagcctgggg ataagtgccc
     5221 tgcggtattg acacttgagg ggcgcgacta ctgacagatg aggggcgcga tccttgacac
     5281 ttgaggggca gagtgctgac agatgagggg cgcacctatt gacatttgag gggctgtcca
     5341 caggcagaaa atccagcatt tgcaagggtt tccgcccgtt tttcggccac cgctaacctg
     5401 tcttttaacc tgcttttaaa ccaatattta taaaccttgt ttttaaccag ggctgcgccc
     5461 tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc
     5521 gctaacgcgg gcctcccatc cccccagggg ctgcgcccct cggccgcgaa cggcctcacc
     5581 ccaaaaatgg cagcgctggc agtccttgcc attgccggga tcggggcagt aacgggatgg
     5641 gcgatcagcc cgaaagctac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5701 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5761 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5821 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5881 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5941 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6001 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6061 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6301 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6601 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6661 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6721 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6781 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6841 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6901 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6961 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7021 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7081 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7141 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7201 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7261 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7321 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7381 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggac
     7441 tctagctaga ggatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct
     7501 ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc
     7561 ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact
     7621 gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg
     7681 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
     7741 acattgcgga cgtttttaat gatcgaatac taacgtctct accagatatc agcttgcatg
     7801 ccggtcgatc tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct
     7861 atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc
     7921 atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca
     7981 gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt
     8041 gccaaatgtt tgaacgatct gcttgactct aggggtcatc agatttcggt gacgggcagg
     8101 accggacggg gcggcaccgg caggctgaag tccagctgcc agaaacccac gtcatgccag
     8161 ttcccgtgct tgaagccggc cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg
     8221 tgcatgcgca cgctcgggtc gttgggcagc ccgatgacag cgaccacgct cttgaagccc
     8281 tgtgcctcca gggacttcag caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg
     8341 tggcgggggg agacgtacac ggtcgactcg gccgtccagt cgtaggcgtt gcgtgccttc
     8401 cagggacccg cgtaggcgat gccggcgacc tcgccgtcca cctcggcgac gagccaggga
     8461 tagcgctccc gcagacggac gaggtcgtcc gtccactcct gcggttcctg cggctcggta
     8521 cggaagttga ccgtgcttgt ctcgatgtag tggttgacga tggtgcagac cgccggcatg
     8581 tccgcctcgg tggcacggcg gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc
     8641 ctcgatcgag ttgagagtga atatgagact ctaattggat accgagggga atttatggaa
     8701 cgtcagtgga gcatttttga caagaaatat ttgctagctg atagtgacct taggcgactt
     8761 ttgaacgcgc aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg
     8821 cggctcagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc
     8881 ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctccgctcat gatcagattg
     8941 tcgtttcccg ccttcggttt gggcgcgccc ggttcagaag accagagggc tattgagact
     9001 tttcaacaaa gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac
     9061 ttcatcgaaa ggacagtaga aaaggaagat ggcttctaca aatgccatca ttgcgataaa
     9121 ggaaaggcta tcgttcaaga tgcctctacc gacagtggtc ccaaagatgg acccccaccc
     9181 acgaggaaca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga
     9241 tgtgatacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
     9301 cagaagacca gagggctat
//
