LOCUS       sequence-169564-        8185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cf-4 split YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8185)
  AUTHORS   Postma J, Liebrand TW, Bi G, Evrard A, Bye RR, Mbengue M, Kuhn H, 
            Joosten MH, Robatzek S
  TITLE     Avr4 promotes Cf-4 receptor-like protein association with the 
            BAK1/SERK3 receptor-like kinase to initiate receptor endocytosis and
            plant immunity.
  JOURNAL   New Phytol. 2016 Apr;210(2):627-42. doi: 10.1111/nph.13802. Epub 
            2016 Jan 14.
  PUBMED    26765243
REFERENCE   2  (bases 1 to 8185)
  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: "10.1111/nph.13802";
            journalName: "New Phytol"; date: "2016-04"; volume: "210"; issue: 
            "2"; pages: "627-42"
COMMENT     Sequence Label: p35S::Cf-4-YFPn
FEATURES             Location/Qualifiers
     source          1..8185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        535..626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(627..695,696..1487)
                     /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(845..864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1658..2246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2147..2166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2558..2582
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(2676..2928)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2948..3499)
                     /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(3541..3720)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        4189..4533
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4477..4497
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     4535..4551
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4536..4552
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4568..4592
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7014..7038)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(7046..7048,7049..7051,7052..7510)
                     /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(7091..7112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7352..7371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    7802..7826
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      join(7938..8185,1..462)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat
       61 gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac
      121 tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc
      181 gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt
      241 ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat
      301 aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg
      361 tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc ccccaggggc
      421 tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agcgctggca gtccttgcca
      481 ttgccgggat cggggcagta acgggatggg cgatcagccc gaaagctacc cctatttgtt
      541 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      601 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      661 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      721 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      781 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      841 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      901 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
      961 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1021 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1081 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1141 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1201 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1261 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1321 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1381 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1441 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1501 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1561 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1801 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2281 ccttttgctg gccttttgct cacatggact ctagctagag gatcacaggc agcaacgctc
     2341 tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca
     2401 gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa
     2461 cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg
     2521 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa
     2581 acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg atcgaatact
     2641 aacgtctcta ccagatatca gcttgcatgc cggtcgatct agtaacatag atgacaccgc
     2701 gcgcgataat ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt attaaatgta
     2761 taattgcggg actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt
     2821 aattattaca tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc
     2881 aacaggattc aatcttaaga aactttattg ccaaatgttt gaacgatctg cttgactcta
     2941 ggggtcatca gatttcggtg acgggcagga ccggacgggg cggcaccggc aggctgaagt
     3001 ccagctgcca gaaacccacg tcatgccagt tcccgtgctt gaagccggcc gcccgcagca
     3061 tgccgcgggg ggcatatccg agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc
     3121 cgatgacagc gaccacgctc ttgaagccct gtgcctccag ggacttcagc aggtgggtgt
     3181 agagcgtgga gcccagtccc gtccgctggt ggcgggggga gacgtacacg gtcgactcgg
     3241 ccgtccagtc gtaggcgttg cgtgccttcc agggacccgc gtaggcgatg ccggcgacct
     3301 cgccgtccac ctcggcgacg agccagggat agcgctcccg cagacggacg aggtcgtccg
     3361 tccactcctg cggttcctgc ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt
     3421 ggttgacgat ggtgcagacc gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg
     3481 ggcgtcgttc tgggctcatg gtagatcccc tcgatcgagt tgagagtgaa tatgagactc
     3541 taattggata ccgaggggaa tttatggaac gtcagtggag catttttgac aagaaatatt
     3601 tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt ctgacgtatg
     3661 tgcttagctc attaaactcc agaaacccgc ggctcagtgg ctccttcaac gttgcggttc
     3721 tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact
     3781 cccttaattc tccgctcatg atcagattgt cgtttcccgc cttcggtttg ggcgcgcccg
     3841 gttcagaaga ccagagggct attgagactt ttcaacaaag ggtaatatcg ggaaacctcc
     3901 tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa aaggaagatg
     3961 gcttctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat gcctctaccg
     4021 acagtggtcc caaagatgga cccccaccca cgaggaacat cgtggaaaaa gaagacgttc
     4081 caaccacgtc ttcaaagcaa gtggattgat gtgatacatg gtggagcacg acactctcgt
     4141 ctactccaag aatatcaaag atacagtctc agaagaccag agggctattg agacttttca
     4201 acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
     4261 cgaaaggaca gtagaaaagg aagatggctt ctacaaatgc catcattgcg ataaaggaaa
     4321 ggctatcgtt caagatgcct ctaccgacag tggtcccaaa gatggacccc cacccacgag
     4381 gaacatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga
     4441 tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
     4501 tatataagga agttcatttc atttggagag gacctcgagg tcgacggtat cgataagctt
     4561 cccatcaaca agtttgtaca aaaaagcagg ctttatgggt tgtgtaaaac ttgtgttttt
     4621 catgctatat gtctttctct ttcaacttgt ttcctcgtca tccttacctc atttgtgccc
     4681 cgaagatcaa gctcttgctc ttctagaatt caagaacatg tttaccgtta atcctaatgc
     4741 ttctgattat tgttacgaca gaagaactct ttcttggaac aaaagcacaa gttgctgctc
     4801 atgggatggc gttcattgtg acgaaacgac aggacaagtg attgagcttg acctccgttg
     4861 catccaactt caaggcaagt ttcattccaa tagtagcctc tttcaactct ccaatctcaa
     4921 aaggcttgat ttgtcttata atgatttcac tggatcgccc atttcaccta aatttggtga
     4981 gttttcagat ttgacgcatc tcgatttgtc gcattcaagt tttaggggtg taatcccttc
     5041 tgaaatctct catctttcta aactatacgt tcttcgtatt agtctaaatg agcttacttt
     5101 tggtcctcac aattttgaat tgcttcttaa gaacttgacc caattaaaag tgctcgacct
     5161 tgaatctatc aacatctctt ccactattcc tttgaatttc tcttctcatt taacaaatct
     5221 atggcttcca tacacagagt tacgtgggat attgcccgaa agagttttcc acctttccga
     5281 cttagaattt ctcgatttat caagcaatcc ccagctcacg gttaggtttc ccacaaccaa
     5341 atggaatagc agtgcatcac tcatgaagtt atatctctat aatgtgaata ttgatgatag
     5401 gatacctgaa tcatttagcc atctaacttc acttcataag ttgtacatga gtcgttctaa
     5461 tctgtcaggg cctattccta aacctctatg gaatctcacc aacatagtgt ttttggacct
     5521 taataataac catcttgaag gaccaattcc atccaacgta agcggactac gtaacctaca
     5581 aatactttgg ttgtcatcaa acaacttaaa tgggagtata ccatcctgga tattctccct
     5641 tccatcactg atagggttag acttgagcaa taacactttc agtggaaaaa ttcaagagtt
     5701 caagtccaaa acattaagta ccgttactct aaaacaaaat aagctaaaag gtcctattcc
     5761 gaattcactc ctaaaccaga agaacctaca attccttctc ctttcacaca ataatatcag
     5821 tggacatatt tcttcagcta tctgcaatct gaaaacattg atattgttag acttgggaag
     5881 taataatttg gagggaacaa tcccgcaatg cgtggttgag aggaacgaat acctttcgca
     5941 tttggatttg agcaacaaca gacttagtgg gacaatcaat acaactttta gtgttggaaa
     6001 cattttaagg gtcattagct tgcacgggaa taagctaacg gggaaagtcc cacgatctat
     6061 gatcaattgc aagtatttga cactacttga tctaggtaac aatatgttga atgacacatt
     6121 tccaaactgg ttgggatacc tatttcaatt gaagatttta agcttgagat caaataagtt
     6181 gcatggtccc atcaaatctt cagggaatac aaacttgttt atgggtcttc aaattcttga
     6241 tctatcatct aatggattta gtgggaattt acccgaaaga attttgggga atttgcaaac
     6301 catgaaggaa attgatgaga gtacaggatt cccagagtat atttctgatc catatgatat
     6361 ttattacaat tatttgacga caatttctac aaagggacaa gattatgatt ctgttcgaat
     6421 tttggattct aacatgatta tcaatctctc aaagaacaga tttgaaggtc atattccaag
     6481 cattattgga gatcttgttg gacttcgtac gttgaacttg tctcacaatg tcttggaagg
     6541 tcatataccg gcatcatttc aaaatttatc agtactcgaa tctttggatc tctcatctaa
     6601 taaaatcagc ggagaaattc cgcagcagct tgcatccctc acattccttg aagtcttaaa
     6661 tctctctcac aatcatcttg ttggatgcat ccccaaagga aaacaatttg attcgttcgg
     6721 gaacacttcg taccaaggga atgatgggtt acgcggattt ccactctcaa aactttgtgg
     6781 tggtgaagat caagtgacaa ctccagctga gctagatcaa gaagaggagg aagaagattc
     6841 accaatgatc agttggcagg gggttctcgt gggttacggt tgtggacttg ttattggact
     6901 gtccgtaata tacataatgt ggtcaactca atatccagca tggttttcga ggatggattt
     6961 aaagttggaa cacataatta ctacgaaaat gaaaaagcac aagaaaagat ataacccagc
     7021 tttcttgtac aaagtggttg atgggatggt gagcaagggc gaggagctgt tcaccggggt
     7081 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     7141 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     7201 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     7261 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     7321 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     7381 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     7441 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     7501 tatcatggcc taatctagag tccgcaaaaa tcaccagtct ctctctacaa atctatctct
     7561 ctctattttt ctccagaata atgtgtgagt agttcccaga taagggaatt agggttctta
     7621 tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa
     7681 aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtgac cgggcggccg
     7741 ccaccgcggt ggagggggat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt
     7801 ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
     7861 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgta catcaccgac
     7921 gagcaaggca agaccgagcg cctttccgac gctcaccggg ctggttgccc tcgccgctgg
     7981 gctggcggcc gtctatggcc ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag
     8041 acaccggccg ccggcgttgt ggatacctcg cggaaaactt ggccctcact gacagatgag
     8101 gggcggacgt tgacacttga ggggccgact cacccggcgc ggcgttgaca gatgaggggc
     8161 aggctcgatt tcggccggcg acgtg
//
