LOCUS       sequence-176890-        9109 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 9109)
  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 9109)
  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-YFPc
FEATURES             Location/Qualifiers
     source          1..9109
                     /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             3995..4246
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     4183..4204
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4538..4562
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4674..5383
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5456..5547
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5548..5616,5617..6408)
                     /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(5766..5785)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6579..7167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7068..7087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7479..7503
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(7597..7849)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7869..8420)
                     /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(8462..8641)
                     /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 gatggacaag cagaagaacg gcatcaaggc
     4021 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     4081 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     4141 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4201 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat ctagagtccg
     4261 caaaaatcac cagtctctct ctacaaatct atctctctct atttttctcc agaataatgt
     4321 gtgagtagtt cccagataag ggaattaggg ttcttatagg gtttcgctca tgtgttgagc
     4381 atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct
     4441 aattcctaaa accaaaatcc agtgaccggg cggccgccac cgcggtggag ggggatcaga
     4501 ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat tggcgggtaa
     4561 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     4621 tatccgttcg tccatttgta tgtgtacatc accgacgagc aaggcaagac cgagcgcctt
     4681 tccgacgctc accgggctgg ttgccctcgc cgctgggctg gcggccgtct atggccctgc
     4741 aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cggccgccgg cgttgtggat
     4801 acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac acttgagggg
     4861 ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg ccggcgacgt
     4921 ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt ttcccacaga
     4981 tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg ggcgcgacta
     5041 ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac agatgagggg
     5101 cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt tgcaagggtt
     5161 tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa ccaatattta
     5221 taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg ccgaaggggg
     5281 gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc cccccagggg
     5341 ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc agtccttgcc
     5401 attgccggga tcggggcagt aacgggatgg gcgatcagcc cgaaagctac ccctatttgt
     5461 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5521 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5581 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5641 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5701 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5761 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5821 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5881 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5941 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6001 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6061 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6121 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6181 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6241 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6301 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6361 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6421 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6481 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6541 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6601 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6661 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6721 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6781 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6841 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6901 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6961 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7021 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7081 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7141 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7201 gccttttgct ggccttttgc tcacatggac tctagctaga ggatcacagg cagcaacgct
     7261 ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc
     7321 agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca
     7381 acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt
     7441 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
     7501 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gatcgaatac
     7561 taacgtctct accagatatc agcttgcatg ccggtcgatc tagtaacata gatgacaccg
     7621 cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt
     7681 ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt
     7741 taattattac atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg
     7801 caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct
     7861 aggggtcatc agatttcggt gacgggcagg accggacggg gcggcaccgg caggctgaag
     7921 tccagctgcc agaaacccac gtcatgccag ttcccgtgct tgaagccggc cgcccgcagc
     7981 atgccgcggg gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc gttgggcagc
     8041 ccgatgacag cgaccacgct cttgaagccc tgtgcctcca gggacttcag caggtgggtg
     8101 tagagcgtgg agcccagtcc cgtccgctgg tggcgggggg agacgtacac ggtcgactcg
     8161 gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat gccggcgacc
     8221 tcgccgtcca cctcggcgac gagccaggga tagcgctccc gcagacggac gaggtcgtcc
     8281 gtccactcct gcggttcctg cggctcggta cggaagttga ccgtgcttgt ctcgatgtag
     8341 tggttgacga tggtgcagac cgccggcatg tccgcctcgg tggcacggcg gatgtcggcc
     8401 gggcgtcgtt ctgggctcat ggtagatccc ctcgatcgag ttgagagtga atatgagact
     8461 ctaattggat accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat
     8521 ttgctagctg atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat
     8581 gtgcttagct cattaaactc cagaaacccg cggctcagtg gctccttcaa cgttgcggtt
     8641 ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac
     8701 tcccttaatt ctccgctcat gatcagattg tcgtttcccg ccttcggttt gggcgcgccc
     8761 ggttcagaag accagagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc
     8821 ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaagat
     8881 ggcttctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctacc
     8941 gacagtggtc ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa agaagacgtt
     9001 ccaaccacgt cttcaaagca agtggattga tgtgatacat ggtggagcac gacactctcg
     9061 tctactccaa gaatatcaaa gatacagtct cagaagacca gagggctat
//
