LOCUS       sequence-176886-        8989 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 8989)
  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 8989)
  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: ACA8-YFPn
FEATURES             Location/Qualifiers
     source          1..8989
                     /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(3630..3654)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3662..3664,3665..3667,3668..4126)
                     /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(3707..3728)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3968..3987)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4418..4442
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4554..5263
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5336..5427
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5428..5496,5497..6288)
                     /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(5646..5665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6459..7047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6948..6967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7359..7383
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(7477..7729)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7749..8300)
                     /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(8342..8521)
                     /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 ggctacatga cgagtctctt
      421 gaagtcatcg cctggacgac gccgtggagg cgatgtggag tctgggaaga gtgaacacgc
      481 agactctgat agcgacactt tctacatccc atcgaagaat gcttctattg agcggcttca
      541 acagtggaga aaagctgcac tggttctcaa tgcgtcccgg agattccggt acaccttgga
      601 cttgaagaag gagcaagaaa cgagggaaat gagacagaag atcagaagtc atgctcatgc
      661 tcttttggcg gcgaatcgtt ttatggatat gggacgtgag tcaggagttg aaaaaacaac
      721 tgggcctgca actccggctg gtgattttgg aattactcct gaacagcttg tgataatgtc
      781 aaaggatcac aacagtggtg ccctggagca atatggaggg actcaaggat tggccaattt
      841 gctcaagaca aatccagaga aaggtatcag tggcgatgat gatgacttgc taaagcgcaa
      901 gaccatttat ggatcaaaca catatcctcg caagaaaggg aaagggtttc tgaggtttct
      961 ttgggatgct tgccatgatc tcaccttgat cattttaatg gttgctgctg tagcgtcttt
     1021 agcacttggg ataaaaacag agggtatcaa agaaggatgg tatgatggag gaagcattgc
     1081 atttgctgtg attcttgtga ttgttgtgac ggctgtcagt gactacaaac agtcgctcca
     1141 gtttcaaaac ttgaatgatg aaaagagaaa catacatctg gaggtgttaa gaggtggaag
     1201 acgagtggag atttcgatct atgacatagt ggtgggtgat gtcattcctc tcaacattgg
     1261 caatcaggtt cctgctgatg gagtactgat atctggccac tctctcgccc ttgatgaatc
     1321 tagcatgact ggagagagca aaattgttaa caaagacgct aacaaggacc cattcttaat
     1381 gtctggctgt aaagtggcag atggaaatgg ttctatgctg gttactggtg ttggagtcaa
     1441 cactgaatgg ggattgctga tggccagtat ttctgaagac aatggtgaag aaactccctt
     1501 gcaggtgcgc ttaaatgggg tagctacttt tattggttca attggcttag cggtggctgc
     1561 tgctgtgcta gtgattcttc tgactcgata tttcactggt cacactaaag acaataatgg
     1621 aggtcctcaa ttcgttaaag gcaagacaaa agtcggtcat gttattgatg atgtggtcaa
     1681 agtccttacc gtagcggtta caattgtcgt agtggcagtg cctgagggtc ttcctttggc
     1741 tgttacttta acccttgcct attcaatgag gaaaatgatg gcagataagg ctttggtgcg
     1801 gaggctatct gcttgcgaga caatgggctc tgccaccact atttgcagcg ataaaactgg
     1861 aacactaact ctgaatcaga tgactgtggt tgagtcttat gctgggggca agaaaacaga
     1921 tactgaacaa ttgccagcaa ctatcacttc gctagtagtt gaaggaatat ctcaaaacac
     1981 aactggtagt atttttgtcc cagagggtgg aggtgattta gagtactctg gttcaccaac
     2041 agaaaaggcc attcttggct ggggagttaa gctgggaatg aatttcgaga cagcccgatc
     2101 gcagtcttct attcttcatg cttttccatt taactcagag aagaaacgtg gtggtgttgc
     2161 tgtaaaaacg gctgatggtg aagttcatgt tcactggaaa ggagcttctg aaattgtcct
     2221 ggcatcatgc cgaagctaca tcgatgagga tggtaatgtg gcaccaatga ctgacgacaa
     2281 ggcatcgttt ttcaagaatg gcattaatga tatggctgga agaaccttac gatgtgttgc
     2341 actcgccttt agaacctatg aggctgaaaa ggtcccaaca ggcgaagagc tctcaaagtg
     2401 ggtactccca gaggatgatc ttatattatt agctatagtg ggcataaagg atccatgtag
     2461 accaggagtc aaagattcag ttgtgttgtg tcaaaatgct ggtgtcaagg tccgtatggt
     2521 tactggtgac aacgtccaaa ctgccagggc tattgccttg gaatgtggaa tactaagttc
     2581 agatgcagat ctctccgagc ctactctcat tgaaggaaaa tcattccgtg agatgactga
     2641 tgcagaaagg gacaagatta gtgacaaaat atcggttatg ggccgatcat ctcccaatga
     2701 caaacttctg ctggtacaat ctctgagacg acaagggcat gttgttgctg tcaccgggga
     2761 tgggacaaat gatgctcctg cattgcacga ggcagatatt ggtcttgcta tgggaatagc
     2821 aggaacagaa gtggctaagg agagttcgga catcatcatc ttggatgaca actttgcttc
     2881 agttgtcaag gttgttcgat gggggcgatc agtgtatgcc aacattcaga agttcatcca
     2941 atttcagctc acagtcaatg tcgccgctct cgtaattaat gttgtagctg ctatttcgag
     3001 tggtgacgtt ccacttaccg ctgtgcagct tctctgggtt aatctgataa tggacactct
     3061 tggagcattg gctttggcta ctgaaccacc cactgatcac ctcatgggga ggcctccagt
     3121 tggcagaaaa gaacctctta ttaccaacat catgtggaga aacttgctga tccaggctat
     3181 ttatcaagtg agtgtattgt taacactcaa tttccgaggc ataagcatac ttggcctgga
     3241 gcatgaagtg catgaacacg ccactagagt gaagaacaca ataatcttca acgcctttgt
     3301 tctctgccaa gctttcaatg agttcaatgc tcgtaaacca gatgagaaga acatcttcaa
     3361 aggtgtgatc aagaatcgtc tctttatggg aataatagtc ataactcttg tgctccaggt
     3421 tatcattgtt gagttccttg gtaaattcgc ttccacgacg aaactaaact ggaagcaatg
     3481 gcttatctgt gttggcatcg gtgttatcag ttggcctctt gctttggtcg ggaaattcat
     3541 tccggtgcct gcagctccta taagcaacaa attgaaagta ctaaaattct ggggcaagaa
     3601 gaaaaattct tctggagaag gttcactcca cccagctttc ttgtacaaag tggttgatgg
     3661 gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     3721 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     3781 cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     3841 cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     3901 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3961 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4021 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4081 caagctggag tacaactaca acagccacaa cgtctatatc atggcctaat ctagagtccg
     4141 caaaaatcac cagtctctct ctacaaatct atctctctct atttttctcc agaataatgt
     4201 gtgagtagtt cccagataag ggaattaggg ttcttatagg gtttcgctca tgtgttgagc
     4261 atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct
     4321 aattcctaaa accaaaatcc agtgaccggg cggccgccac cgcggtggag ggggatcaga
     4381 ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat tggcgggtaa
     4441 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     4501 tatccgttcg tccatttgta tgtgtacatc accgacgagc aaggcaagac cgagcgcctt
     4561 tccgacgctc accgggctgg ttgccctcgc cgctgggctg gcggccgtct atggccctgc
     4621 aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cggccgccgg cgttgtggat
     4681 acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac acttgagggg
     4741 ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg ccggcgacgt
     4801 ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt ttcccacaga
     4861 tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg ggcgcgacta
     4921 ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac agatgagggg
     4981 cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt tgcaagggtt
     5041 tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa ccaatattta
     5101 taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg ccgaaggggg
     5161 gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc cccccagggg
     5221 ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc agtccttgcc
     5281 attgccggga tcggggcagt aacgggatgg gcgatcagcc cgaaagctac ccctatttgt
     5341 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5401 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5461 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5521 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5581 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5641 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5701 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5761 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5821 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5881 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5941 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6001 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6061 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6121 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6181 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6241 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6301 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6361 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6781 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6841 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6901 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6961 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7021 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7081 gccttttgct ggccttttgc tcacatggac tctagctaga ggatcacagg cagcaacgct
     7141 ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc
     7201 agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca
     7261 acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt
     7321 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
     7381 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gatcgaatac
     7441 taacgtctct accagatatc agcttgcatg ccggtcgatc tagtaacata gatgacaccg
     7501 cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt
     7561 ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt
     7621 taattattac atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg
     7681 caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct
     7741 aggggtcatc agatttcggt gacgggcagg accggacggg gcggcaccgg caggctgaag
     7801 tccagctgcc agaaacccac gtcatgccag ttcccgtgct tgaagccggc cgcccgcagc
     7861 atgccgcggg gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc gttgggcagc
     7921 ccgatgacag cgaccacgct cttgaagccc tgtgcctcca gggacttcag caggtgggtg
     7981 tagagcgtgg agcccagtcc cgtccgctgg tggcgggggg agacgtacac ggtcgactcg
     8041 gccgtccagt cgtaggcgtt gcgtgccttc cagggacccg cgtaggcgat gccggcgacc
     8101 tcgccgtcca cctcggcgac gagccaggga tagcgctccc gcagacggac gaggtcgtcc
     8161 gtccactcct gcggttcctg cggctcggta cggaagttga ccgtgcttgt ctcgatgtag
     8221 tggttgacga tggtgcagac cgccggcatg tccgcctcgg tggcacggcg gatgtcggcc
     8281 gggcgtcgtt ctgggctcat ggtagatccc ctcgatcgag ttgagagtga atatgagact
     8341 ctaattggat accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat
     8401 ttgctagctg atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat
     8461 gtgcttagct cattaaactc cagaaacccg cggctcagtg gctccttcaa cgttgcggtt
     8521 ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac
     8581 tcccttaatt ctccgctcat gatcagattg tcgtttcccg ccttcggttt gggcgcgccc
     8641 ggttcagaag accagagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc
     8701 ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaagat
     8761 ggcttctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctacc
     8821 gacagtggtc ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa agaagacgtt
     8881 ccaaccacgt cttcaaagca agtggattga tgtgatacat ggtggagcac gacactctcg
     8941 tctactccaa gaatatcaaa gatacagtct cagaagacca gagggctat
//
