LOCUS       sequence-176887-        8779 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 8779)
  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 8779)
  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-YFPc
FEATURES             Location/Qualifiers
     source          1..8779
                     /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             3665..3916
                     /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     3853..3874
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4208..4232
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4344..5053
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5126..5217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5218..5286,5287..6078)
                     /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(5436..5455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6249..6837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6738..6757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7149..7173
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(7267..7519)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7539..8090)
                     /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(8132..8311)
                     /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 gatggacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     3721 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3781 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     3841 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3901 ggacgagctg tacaagtaat ctagagtccg caaaaatcac cagtctctct ctacaaatct
     3961 atctctctct atttttctcc agaataatgt gtgagtagtt cccagataag ggaattaggg
     4021 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     4081 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgaccggg
     4141 cggccgccac cgcggtggag ggggatcaga ttgtcgtttc ccgccttcag tttaaactat
     4201 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     4261 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgtacatc
     4321 accgacgagc aaggcaagac cgagcgcctt tccgacgctc accgggctgg ttgccctcgc
     4381 cgctgggctg gcggccgtct atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg
     4441 tgcgagacac cggccgccgg cgttgtggat acctcgcgga aaacttggcc ctcactgaca
     4501 gatgaggggc ggacgttgac acttgagggg ccgactcacc cggcgcggcg ttgacagatg
     4561 aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa
     4621 acgcctgatt ttacgcgagt ttcccacaga tgatgtggac aagcctgggg ataagtgccc
     4681 tgcggtattg acacttgagg ggcgcgacta ctgacagatg aggggcgcga tccttgacac
     4741 ttgaggggca gagtgctgac agatgagggg cgcacctatt gacatttgag gggctgtcca
     4801 caggcagaaa atccagcatt tgcaagggtt tccgcccgtt tttcggccac cgctaacctg
     4861 tcttttaacc tgcttttaaa ccaatattta taaaccttgt ttttaaccag ggctgcgccc
     4921 tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc
     4981 gctaacgcgg gcctcccatc cccccagggg ctgcgcccct cggccgcgaa cggcctcacc
     5041 ccaaaaatgg cagcgctggc agtccttgcc attgccggga tcggggcagt aacgggatgg
     5101 gcgatcagcc cgaaagctac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5161 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5221 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5281 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5341 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5401 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5461 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5521 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5581 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5641 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5701 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5761 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5821 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5881 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5941 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6001 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6061 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6361 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggac
     6901 tctagctaga ggatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct
     6961 ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc
     7021 ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact
     7081 gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg
     7141 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
     7201 acattgcgga cgtttttaat gatcgaatac taacgtctct accagatatc agcttgcatg
     7261 ccggtcgatc tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct
     7321 atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc
     7381 atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca
     7441 gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt
     7501 gccaaatgtt tgaacgatct gcttgactct aggggtcatc agatttcggt gacgggcagg
     7561 accggacggg gcggcaccgg caggctgaag tccagctgcc agaaacccac gtcatgccag
     7621 ttcccgtgct tgaagccggc cgcccgcagc atgccgcggg gggcatatcc gagcgcctcg
     7681 tgcatgcgca cgctcgggtc gttgggcagc ccgatgacag cgaccacgct cttgaagccc
     7741 tgtgcctcca gggacttcag caggtgggtg tagagcgtgg agcccagtcc cgtccgctgg
     7801 tggcgggggg agacgtacac ggtcgactcg gccgtccagt cgtaggcgtt gcgtgccttc
     7861 cagggacccg cgtaggcgat gccggcgacc tcgccgtcca cctcggcgac gagccaggga
     7921 tagcgctccc gcagacggac gaggtcgtcc gtccactcct gcggttcctg cggctcggta
     7981 cggaagttga ccgtgcttgt ctcgatgtag tggttgacga tggtgcagac cgccggcatg
     8041 tccgcctcgg tggcacggcg gatgtcggcc gggcgtcgtt ctgggctcat ggtagatccc
     8101 ctcgatcgag ttgagagtga atatgagact ctaattggat accgagggga atttatggaa
     8161 cgtcagtgga gcatttttga caagaaatat ttgctagctg atagtgacct taggcgactt
     8221 ttgaacgcgc aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg
     8281 cggctcagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc
     8341 ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctccgctcat gatcagattg
     8401 tcgtttcccg ccttcggttt gggcgcgccc ggttcagaag accagagggc tattgagact
     8461 tttcaacaaa gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac
     8521 ttcatcgaaa ggacagtaga aaaggaagat ggcttctaca aatgccatca ttgcgataaa
     8581 ggaaaggcta tcgttcaaga tgcctctacc gacagtggtc ccaaagatgg acccccaccc
     8641 acgaggaaca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga
     8701 tgtgatacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
     8761 cagaagacca gagggctat
//
