LOCUS       sequence-176889-        9263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent labelling of ACA8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9263)
  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 9263)
  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-mYFP
FEATURES             Location/Qualifiers
     source          1..9263
                     /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    398..422
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3648..3672)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3680..3682,3683..3685,3686..4399)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3725..3746)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3986..4005)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4333..4354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4692..4716
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4828..5537
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5610..5701
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5702..5770,5771..6562)
                     /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(5920..5939)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6733..7321
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7222..7241
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7633..7657
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(7751..8003)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8023..8574)
                     /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(8616..8795)
                     /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 ttgatatcga attcctgcag cccatcaaca agtttgtaca aaaaagcagg
      421 ctacatgacg agtctcttga agtcatcgcc tggacgacgc cgtggaggcg atgtggagtc
      481 tgggaagagt gaacacgcag actctgatag cgacactttc tacatcccat cgaagaatgc
      541 ttctattgag cggcttcaac agtggagaaa agctgcactg gttctcaatg cgtcccggag
      601 attccggtac accttggact tgaagaagga gcaagaaacg agggaaatga gacagaagat
      661 cagaagtcat gctcatgctc ttttggcggc gaatcgtttt atggatatgg gacgtgagtc
      721 aggagttgaa aaaacaactg ggcctgcaac tccggctggt gattttggaa ttactcctga
      781 acagcttgtg ataatgtcaa aggatcacaa cagtggtgcc ctggagcaat atggagggac
      841 tcaaggattg gccaatttgc tcaagacaaa tccagagaaa ggtatcagtg gcgatgatga
      901 tgacttgcta aagcgcaaga ccatttatgg atcaaacaca tatcctcgca agaaagggaa
      961 agggtttctg aggtttcttt gggatgcttg ccatgatctc accttgatca ttttaatggt
     1021 tgctgctgta gcgtctttag cacttgggat aaaaacagag ggtatcaaag aaggatggta
     1081 tgatggagga agcattgcat ttgctgtgat tcttgtgatt gttgtgacgg ctgtcagtga
     1141 ctacaaacag tcgctccagt ttcaaaactt gaatgatgaa aagagaaaca tacatctgga
     1201 ggtgttaaga ggtggaagac gagtggagat ttcgatctat gacatagtgg tgggtgatgt
     1261 cattcctctc aacattggca atcaggttcc tgctgatgga gtactgatat ctggccactc
     1321 tctcgccctt gatgaatcta gcatgactgg agagagcaaa attgttaaca aagacgctaa
     1381 caaggaccca ttcttaatgt ctggctgtaa agtggcagat ggaaatggtt ctatgctggt
     1441 tactggtgtt ggagtcaaca ctgaatgggg attgctgatg gccagtattt ctgaagacaa
     1501 tggtgaagaa actcccttgc aggtgcgctt aaatggggta gctactttta ttggttcaat
     1561 tggcttagcg gtggctgctg ctgtgctagt gattcttctg actcgatatt tcactggtca
     1621 cactaaagac aataatggag gtcctcaatt cgttaaaggc aagacaaaag tcggtcatgt
     1681 tattgatgat gtggtcaaag tccttaccgt agcggttaca attgtcgtag tggcagtgcc
     1741 tgagggtctt cctttggctg ttactttaac ccttgcctat tcaatgagga aaatgatggc
     1801 agataaggct ttggtgcgga ggctatctgc ttgcgagaca atgggctctg ccaccactat
     1861 ttgcagcgat aaaactggaa cactaactct gaatcagatg actgtggttg agtcttatgc
     1921 tgggggcaag aaaacagata ctgaacaatt gccagcaact atcacttcgc tagtagttga
     1981 aggaatatct caaaacacaa ctggtagtat ttttgtccca gagggtggag gtgatttaga
     2041 gtactctggt tcaccaacag aaaaggccat tcttggctgg ggagttaagc tgggaatgaa
     2101 tttcgagaca gcccgatcgc agtcttctat tcttcatgct tttccattta actcagagaa
     2161 gaaacgtggt ggtgttgctg taaaaacggc tgatggtgaa gttcatgttc actggaaagg
     2221 agcttctgaa attgtcctgg catcatgccg aagctacatc gatgaggatg gtaatgtggc
     2281 accaatgact gacgacaagg catcgttttt caagaatggc attaatgata tggctggaag
     2341 aaccttacga tgtgttgcac tcgcctttag aacctatgag gctgaaaagg tcccaacagg
     2401 cgaagagctc tcaaagtggg tactcccaga ggatgatctt atattattag ctatagtggg
     2461 cataaaggat ccatgtagac caggagtcaa agattcagtt gtgttgtgtc aaaatgctgg
     2521 tgtcaaggtc cgtatggtta ctggtgacaa cgtccaaact gccagggcta ttgccttgga
     2581 atgtggaata ctaagttcag atgcagatct ctccgagcct actctcattg aaggaaaatc
     2641 attccgtgag atgactgatg cagaaaggga caagattagt gacaaaatat cggttatggg
     2701 ccgatcatct cccaatgaca aacttctgct ggtacaatct ctgagacgac aagggcatgt
     2761 tgttgctgtc accggggatg ggacaaatga tgctcctgca ttgcacgagg cagatattgg
     2821 tcttgctatg ggaatagcag gaacagaagt ggctaaggag agttcggaca tcatcatctt
     2881 ggatgacaac tttgcttcag ttgtcaaggt tgttcgatgg gggcgatcag tgtatgccaa
     2941 cattcagaag ttcatccaat ttcagctcac agtcaatgtc gccgctctcg taattaatgt
     3001 tgtagctgct atttcgagtg gtgacgttcc acttaccgct gtgcagcttc tctgggttaa
     3061 tctgataatg gacactcttg gagcattggc tttggctact gaaccaccca ctgatcacct
     3121 catggggagg cctccagttg gcagaaaaga acctcttatt accaacatca tgtggagaaa
     3181 cttgctgatc caggctattt atcaagtgag tgtattgtta acactcaatt tccgaggcat
     3241 aagcatactt ggcctggagc atgaagtgca tgaacacgcc actagagtga agaacacaat
     3301 aatcttcaac gcctttgttc tctgccaagc tttcaatgag ttcaatgctc gtaaaccaga
     3361 tgagaagaac atcttcaaag gtgtgatcaa gaatcgtctc tttatgggaa taatagtcat
     3421 aactcttgtg ctccaggtta tcattgttga gttccttggt aaattcgctt ccacgacgaa
     3481 actaaactgg aagcaatggc ttatctgtgt tggcatcggt gttatcagtt ggcctcttgc
     3541 tttggtcggg aaattcattc cggtgcctgc agctcctata agcaacaaat tgaaagtact
     3601 aaaattctgg ggcaagaaga aaaattcttc tggagaaggt tcactccacc cagctttctt
     3661 gtacaaagtg gttgatggga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3721 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3781 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3841 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgaagt gcttcgcccg
     3901 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3961 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4021 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4081 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4141 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4201 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4261 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     4321 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat
     4381 ggacgagctg tacaagtaag agctctagag tccgcaaaaa tcaccagtct ctctctacaa
     4441 atctatctct ctctattttt ctccagaata atgtgtgagt agttcccaga taagggaatt
     4501 agggttctta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt
     4561 gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtgac
     4621 cgggcggccg ccaccgcggt ggagggggat cagattgtcg tttcccgcct tcagtttaaa
     4681 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     4741 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgta
     4801 catcaccgac gagcaaggca agaccgagcg cctttccgac gctcaccggg ctggttgccc
     4861 tcgccgctgg gctggcggcc gtctatggcc ctgcaaacgc gccagaaacg ccgtcgaagc
     4921 cgtgtgcgag acaccggccg ccggcgttgt ggatacctcg cggaaaactt ggccctcact
     4981 gacagatgag gggcggacgt tgacacttga ggggccgact cacccggcgc ggcgttgaca
     5041 gatgaggggc aggctcgatt tcggccggcg acgtggagct ggccagcctc gcaaatcggc
     5101 gaaaacgcct gattttacgc gagtttccca cagatgatgt ggacaagcct ggggataagt
     5161 gccctgcggt attgacactt gaggggcgcg actactgaca gatgaggggc gcgatccttg
     5221 acacttgagg ggcagagtgc tgacagatga ggggcgcacc tattgacatt tgaggggctg
     5281 tccacaggca gaaaatccag catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa
     5341 cctgtctttt aacctgcttt taaaccaata tttataaacc ttgtttttaa ccagggctgc
     5401 gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc ccccttctcg aaccctcccg
     5461 gcccgctaac gcgggcctcc catcccccca ggggctgcgc ccctcggccg cgaacggcct
     5521 caccccaaaa atggcagcgc tggcagtcct tgccattgcc gggatcgggg cagtaacggg
     5581 atgggcgatc agcccgaaag ctacccctat ttgtttattt ttctaaatac attcaaatat
     5641 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5701 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5761 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5821 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5881 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5941 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6001 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6061 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6121 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6181 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6241 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6301 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6361 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6421 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6481 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6541 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6601 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6661 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6721 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6781 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6841 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6901 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6961 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7021 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7081 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7141 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7201 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7261 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7321 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7381 ggactctagc tagaggatca caggcagcaa cgctctgtca tcgttacaat caacatgcta
     7441 ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag
     7501 catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg
     7561 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
     7621 ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa
     7681 taacacattg cggacgtttt taatgatcga atactaacgt ctctaccaga tatcagcttg
     7741 catgccggtc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg
     7801 cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa
     7861 acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc
     7921 aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt
     7981 tattgccaaa tgtttgaacg atctgcttga ctctaggggt catcagattt cggtgacggg
     8041 caggaccgga cggggcggca ccggcaggct gaagtccagc tgccagaaac ccacgtcatg
     8101 ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc
     8161 ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa
     8221 gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg
     8281 ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg cgttgcgtgc
     8341 cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca
     8401 gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc
     8461 ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg
     8521 catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcatggtaga
     8581 tcccctcgat cgagttgaga gtgaatatga gactctaatt ggataccgag gggaatttat
     8641 ggaacgtcag tggagcattt ttgacaagaa atatttgcta gctgatagtg accttaggcg
     8701 acttttgaac gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa
     8761 cccgcggctc agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct
     8821 tgtcccgcgt catcggcggg ggtcataacg tgactccctt aattctccgc tcatgatcag
     8881 attgtcgttt cccgccttcg gtttgggcgc gcccggttca gaagaccaga gggctattga
     8941 gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg
     9001 tcacttcatc gaaaggacag tagaaaagga agatggcttc tacaaatgcc atcattgcga
     9061 taaaggaaag gctatcgttc aagatgcctc taccgacagt ggtcccaaag atggaccccc
     9121 acccacgagg aacatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
     9181 ttgatgtgat acatggtgga gcacgacact ctcgtctact ccaagaatat caaagataca
     9241 gtctcagaag accagagggc tat
//
