LOCUS       sequence-169540-        7975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cf-4 slplit YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  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 7975)
  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-YFPc
FEATURES             Location/Qualifiers
     source          1..7975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      537..1125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1026..1045
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1437..1461
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(1555..1807)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1827..2378)
                     /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(2420..2599)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        3068..3412
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3356..3376
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     3414..3430
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3415..3431
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3447..3471
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5893..5917)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5928..6179
                     /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     6116..6137
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6471..6495
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      6607..7316
                     /label=oriV
                     /note="incP origin of replication"
     promoter        7389..7480
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7481..7549,7550..7975,1..366)
                     /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(7699..7718)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
       61 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      121 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      181 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      241 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      301 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
      421 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
      481 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
      541 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      601 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
      661 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
      721 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      781 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      841 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      901 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      961 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1021 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1081 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1141 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggactc tagctagagg
     1201 atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc gcgagatcat
     1261 ccgtgtttca aacccggcag cttagttgcc gttcttccga atagcatcgg taacatgagc
     1321 aaagtctgcc gccttacaac ggctctcccg ctgacgccgt cccggactga tgggctgcct
     1381 gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc
     1441 aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg
     1501 tttttaatga tcgaatacta acgtctctac cagatatcag cttgcatgcc ggtcgatcta
     1561 gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt
     1621 ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata
     1681 acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga
     1741 taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg
     1801 aacgatctgc ttgactctag gggtcatcag atttcggtga cgggcaggac cggacggggc
     1861 ggcaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt cccgtgcttg
     1921 aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg catgcgcacg
     1981 ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg tgcctccagg
     2041 gacttcagca ggtgggtgta gagcgtggag cccagtcccg tccgctggtg gcggggggag
     2101 acgtacacgg tcgactcggc cgtccagtcg taggcgttgc gtgccttcca gggacccgcg
     2161 taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata gcgctcccgc
     2221 agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg gaagttgacc
     2281 gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg
     2341 gcacggcgga tgtcggccgg gcgtcgttct gggctcatgg tagatcccct cgatcgagtt
     2401 gagagtgaat atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc
     2461 atttttgaca agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa
     2521 taatggtttc tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctcagtggc
     2581 tccttcaacg ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg
     2641 cgggggtcat aacgtgactc ccttaattct ccgctcatga tcagattgtc gtttcccgcc
     2701 ttcggtttgg gcgcgcccgg ttcagaagac cagagggcta ttgagacttt tcaacaaagg
     2761 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcgaaagg
     2821 acagtagaaa aggaagatgg cttctacaaa tgccatcatt gcgataaagg aaaggctatc
     2881 gttcaagatg cctctaccga cagtggtccc aaagatggac ccccacccac gaggaacatc
     2941 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatacatgg
     3001 tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaga
     3061 gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc
     3121 cagctatctg tcacttcatc gaaaggacag tagaaaagga agatggcttc tacaaatgcc
     3181 atcattgcga taaaggaaag gctatcgttc aagatgcctc taccgacagt ggtcccaaag
     3241 atggaccccc acccacgagg aacatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     3301 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
     3361 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acctcgaggt
     3421 cgacggtatc gataagcttc ccatcaacaa gtttgtacaa aaaagcaggc tttatgggtt
     3481 gtgtaaaact tgtgtttttc atgctatatg tctttctctt tcaacttgtt tcctcgtcat
     3541 ccttacctca tttgtgcccc gaagatcaag ctcttgctct tctagaattc aagaacatgt
     3601 ttaccgttaa tcctaatgct tctgattatt gttacgacag aagaactctt tcttggaaca
     3661 aaagcacaag ttgctgctca tgggatggcg ttcattgtga cgaaacgaca ggacaagtga
     3721 ttgagcttga cctccgttgc atccaacttc aaggcaagtt tcattccaat agtagcctct
     3781 ttcaactctc caatctcaaa aggcttgatt tgtcttataa tgatttcact ggatcgccca
     3841 tttcacctaa atttggtgag ttttcagatt tgacgcatct cgatttgtcg cattcaagtt
     3901 ttaggggtgt aatcccttct gaaatctctc atctttctaa actatacgtt cttcgtatta
     3961 gtctaaatga gcttactttt ggtcctcaca attttgaatt gcttcttaag aacttgaccc
     4021 aattaaaagt gctcgacctt gaatctatca acatctcttc cactattcct ttgaatttct
     4081 cttctcattt aacaaatcta tggcttccat acacagagtt acgtgggata ttgcccgaaa
     4141 gagttttcca cctttccgac ttagaatttc tcgatttatc aagcaatccc cagctcacgg
     4201 ttaggtttcc cacaaccaaa tggaatagca gtgcatcact catgaagtta tatctctata
     4261 atgtgaatat tgatgatagg atacctgaat catttagcca tctaacttca cttcataagt
     4321 tgtacatgag tcgttctaat ctgtcagggc ctattcctaa acctctatgg aatctcacca
     4381 acatagtgtt tttggacctt aataataacc atcttgaagg accaattcca tccaacgtaa
     4441 gcggactacg taacctacaa atactttggt tgtcatcaaa caacttaaat gggagtatac
     4501 catcctggat attctccctt ccatcactga tagggttaga cttgagcaat aacactttca
     4561 gtggaaaaat tcaagagttc aagtccaaaa cattaagtac cgttactcta aaacaaaata
     4621 agctaaaagg tcctattccg aattcactcc taaaccagaa gaacctacaa ttccttctcc
     4681 tttcacacaa taatatcagt ggacatattt cttcagctat ctgcaatctg aaaacattga
     4741 tattgttaga cttgggaagt aataatttgg agggaacaat cccgcaatgc gtggttgaga
     4801 ggaacgaata cctttcgcat ttggatttga gcaacaacag acttagtggg acaatcaata
     4861 caacttttag tgttggaaac attttaaggg tcattagctt gcacgggaat aagctaacgg
     4921 ggaaagtccc acgatctatg atcaattgca agtatttgac actacttgat ctaggtaaca
     4981 atatgttgaa tgacacattt ccaaactggt tgggatacct atttcaattg aagattttaa
     5041 gcttgagatc aaataagttg catggtccca tcaaatcttc agggaataca aacttgttta
     5101 tgggtcttca aattcttgat ctatcatcta atggatttag tgggaattta cccgaaagaa
     5161 ttttggggaa tttgcaaacc atgaaggaaa ttgatgagag tacaggattc ccagagtata
     5221 tttctgatcc atatgatatt tattacaatt atttgacgac aatttctaca aagggacaag
     5281 attatgattc tgttcgaatt ttggattcta acatgattat caatctctca aagaacagat
     5341 ttgaaggtca tattccaagc attattggag atcttgttgg acttcgtacg ttgaacttgt
     5401 ctcacaatgt cttggaaggt catataccgg catcatttca aaatttatca gtactcgaat
     5461 ctttggatct ctcatctaat aaaatcagcg gagaaattcc gcagcagctt gcatccctca
     5521 cattccttga agtcttaaat ctctctcaca atcatcttgt tggatgcatc cccaaaggaa
     5581 aacaatttga ttcgttcggg aacacttcgt accaagggaa tgatgggtta cgcggatttc
     5641 cactctcaaa actttgtggt ggtgaagatc aagtgacaac tccagctgag ctagatcaag
     5701 aagaggagga agaagattca ccaatgatca gttggcaggg ggttctcgtg ggttacggtt
     5761 gtggacttgt tattggactg tccgtaatat acataatgtg gtcaactcaa tatccagcat
     5821 ggttttcgag gatggattta aagttggaac acataattac tacgaaaatg aaaaagcaca
     5881 agaaaagata taacccagct ttcttgtaca aagtggttga tgggatggac aagcagaaga
     5941 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     6001 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     6061 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     6121 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     6181 aatctagagt ccgcaaaaat caccagtctc tctctacaaa tctatctctc tctatttttc
     6241 tccagaataa tgtgtgagta gttcccagat aagggaatta gggttcttat agggtttcgc
     6301 tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat
     6361 caataaaatt tctaattcct aaaaccaaaa tccagtgacc gggcggccgc caccgcggtg
     6421 gagggggatc agattgtcgt ttcccgcctt cagtttaaac tatcagtgtt tgacaggata
     6481 tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg
     6541 gcgtgaaaag gtttatccgt tcgtccattt gtatgtgtac atcaccgacg agcaaggcaa
     6601 gaccgagcgc ctttccgacg ctcaccgggc tggttgccct cgccgctggg ctggcggccg
     6661 tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga caccggccgc
     6721 cggcgttgtg gatacctcgc ggaaaacttg gccctcactg acagatgagg ggcggacgtt
     6781 gacacttgag gggccgactc acccggcgcg gcgttgacag atgaggggca ggctcgattt
     6841 cggccggcga cgtggagctg gccagcctcg caaatcggcg aaaacgcctg attttacgcg
     6901 agtttcccac agatgatgtg gacaagcctg gggataagtg ccctgcggta ttgacacttg
     6961 aggggcgcga ctactgacag atgaggggcg cgatccttga cacttgaggg gcagagtgct
     7021 gacagatgag gggcgcacct attgacattt gaggggctgt ccacaggcag aaaatccagc
     7081 atttgcaagg gtttccgccc gtttttcggc caccgctaac ctgtctttta acctgctttt
     7141 aaaccaatat ttataaacct tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc
     7201 acgccgaagg ggggtgcccc cccttctcga accctcccgg cccgctaacg cgggcctccc
     7261 atccccccag gggctgcgcc cctcggccgc gaacggcctc accccaaaaa tggcagcgct
     7321 ggcagtcctt gccattgccg ggatcggggc agtaacggga tgggcgatca gcccgaaagc
     7381 tacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7441 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7501 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7561 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7621 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7681 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7741 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7801 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7861 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7921 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccg
//
