LOCUS       sequence-288214-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector FLS2 FLS2_pECIA14 should be used with bait vector FLS2 
            FLS2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  AUTHORS   Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, 
            Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger 
            N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, 
            Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir 
            Y
  TITLE     An extracellular network of Arabidopsis leucine-rich repeat receptor
            kinases.
  JOURNAL   Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. 
            Epub 2018 Jan 10.
  PUBMED    29320478
REFERENCE   2  (bases 1 to 7568)
  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: 
            "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 
            18"; volume: "553"; issue: "7688"; pages: "342-346"
COMMENT     Sequence Label: FLS2 FLS2_pECIA14
FEATURES             Location/Qualifiers
     source          1..7568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        23..449
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     467..520
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2864..2887
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4562..4585
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4601..4618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4641..4658)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4691..4825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4715..4734)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4769..4788
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4969..4985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4969..4985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4982..5004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4993..5009
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5017..5023,5024..5041,5042..5047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5062..5083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5200..5217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5371..5959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5451..5470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6130..6921,6922..6990))
                     /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     6753..6772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6991..7095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7163..7181
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7219..7241)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7341..7360
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7554..7568,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
       61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
      121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
      181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
      241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
      301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
      361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
      421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
      481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg aagcagtcct tcgagcccga
      541 gatcgaggct ctgaagtcgt tcaagaacgg tatctccaac gaccccctgg gtgtcctgtc
      601 cgactggacc atcatcggtt ccctgcgtca ctgcaactgg accggtatca cctgcgactc
      661 caccggtcac gtggtgtccg tgtccctgct ggagaagcag ctggagggcg tgctgtcccc
      721 cgctatcgct aacctgacct acctgcaggt gctggacctg acctccaact ccttcaccgg
      781 caagatcccc gctgagatcg gcaagctgac cgagctgaac cagctgatcc tgtacctgaa
      841 ctacttctcc ggttccatcc cctccggtat ctgggagctg aagaacatct tctacctgga
      901 cctgcgtaac aacctgctgt ccggcgacgt gcccgaggaa atctgcaaga cctcctccct
      961 ggtgctgatc ggtttcgact acaacaacct gaccggaaag atccctgagt gcctgggtga
     1021 cctggtgcac ctgcagatgt tcgtggctgc tggcaaccac ctgactggct ccatccccgt
     1081 gtccatcggc accctggcta acctcaccga cctggacctc tccggcaacc agctcactgg
     1141 aaagatcccc cgtgacttcg gcaacctgct gaacctgcag tctctggtcc tgaccgagaa
     1201 cctgctggag ggcgacatcc ctgccgagat cggaaactgc tcctctctgg tgcagctgga
     1261 gctgtacgac aaccagctga ccggcaagat cccagccgag ctgggcaacc tggtccagct
     1321 gcaggctctg cgtatctaca agaacaagct gacctcctcc atcccttcct ccctgttccg
     1381 tctgacccag ctgacccacc tgggactgtc cgagaaccac ctcgtgggtc ccatctctga
     1441 ggaaatcggt ttcctggagt ccctggaggt gctgaccctg cactctaaca acttcaccgg
     1501 cgagttccct cagtccatca ccaacctgag gaacctgacc gtgctgaccg tcggtttcaa
     1561 caacatctct ggcgagctgc ccgctgacct gggtctgctg accaacctgc gcaacctgtc
     1621 cgctcacgac aacctcctca ccggtcccat tccatcctcc atctccaact gcaccggcct
     1681 gaagctgctc gacctgtccc acaaccagat gactggagaa atcccccgtg gtttcggtcg
     1741 tatgaacctg acattcatct ccatcggtcg taaccacttc actggcgaga tccctgacga
     1801 catcttcaac tgctccaacc tggagactct gtccgtggct gacaacaacc tcactggcac
     1861 cctgaagccc ctgatcggaa agctgcagaa gctgcgtatc ctgcaggtgt cctacaactc
     1921 cctgaccggt cctatccctc gcgagatcgg caacctcaag gacctgaaca tcctgtacct
     1981 ccactccaac ggtttcaccg gtcgtattcc ccgcgagatg tccaacctga ccctgctcca
     2041 gggcctgcgc atgtactcta acgacctgga gggccccatc ccagaggaaa tgttcgacat
     2101 gaagctgctg tccgtcctgg acctcagcaa caacaagttc tccggccaga tccctgctct
     2161 gttctccaag ctggagtctc tcacctacct gtctctgcag ggcaacaagt tcaacggttc
     2221 tatccccgct tccctcaagt ccctgtctct gctgaacacc ttcgacatct ccgacaacct
     2281 gctgactggt actatccccg gcgagctgct ggcttccctg aagaacatgc agctgtacct
     2341 caacttctcc aacaacctgc tcaccggaac catccccaag gaactgggaa agctggaaat
     2401 ggtgcaggaa atcgacctgt ctaacaacct cttctccggt agcatccctc gttcattgca
     2461 agcctgtaag aacgtgttca ccctggactt ctcccagaac aacctgtctg gtcacatccc
     2521 cgacgaggtg ttccagggca tggacatgat catctccctg aacctgtccc gtaactcctt
     2581 cagcggagag atcccacagt ccttcggcaa catgactcac ctggtgtctc tggacctgtc
     2641 ctctaacaac ctgaccggcg agatccccga gtctctggct aacctgtcca ctctgaagca
     2701 cctgaagctg gcttccaaca acctcaaggg tcacgtgccc gagtccggcg tgttcaagaa
     2761 catcaacgcc tccgacctga tgggcaacac cgacctgtgc ggttccaaga agcctctgaa
     2821 gccctgcacc atcaagcaga agtcctccca cttctccaag cgcctggagg tgctgttcca
     2881 gggaccaagc ggcaatcccg gcggcgatct ggccccccag atgctgcgcg agctgcagga
     2941 gaccaatgcc gccctgcagg acgtgcgcga gctgctgcgc caccgcgtga aggagatcac
     3001 cttcctgaag aacaccgtga tggagtgcga cgcctgcggc atgcagccct ctagaggctc
     3061 cggctccatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     3121 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     3181 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     3241 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     3301 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     3361 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     3421 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     3481 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     3541 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     3601 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     3661 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     3721 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     3781 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     3841 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     3901 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     3961 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     4021 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     4081 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     4141 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     4201 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     4261 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     4321 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     4381 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     4441 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     4501 ccccgccggc accaccgacg ccgcgcaccc gggttctaga ggaagcacct ccggcgataa
     4561 cgattacaag gatgatgatg ataaggccag cggcaccggt catcatcacc atcaccattg
     4621 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     4681 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4741 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4801 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4861 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     4921 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     4981 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5041 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5101 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5161 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5221 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5281 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5341 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5401 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5461 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5521 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5581 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5641 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5701 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5761 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5821 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5881 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5941 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6001 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6061 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6121 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6181 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6241 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6301 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6361 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6421 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6481 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6541 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6601 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6661 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6721 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6781 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6841 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6901 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6961 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7021 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7081 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7141 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     7201 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     7261 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7321 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     7381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7441 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7501 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7561 acgacgtt
//
