LOCUS       sequence-296419-        6956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RLK7/KUK2 C12_pECIA14 should be used with bait vector 
            RLK7/KUK2 C12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  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 6956)
  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: RLK7/KUK2 C12_pECIA14
FEATURES             Location/Qualifiers
     source          1..6956
                     /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             2252..2275
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3950..3973
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3989..4006
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4029..4046)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4079..4213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4103..4122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4157..4176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4357..4373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4357..4373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4370..4392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4381..4397
                     /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(4405..4411,4412..4429,4430..4435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4450..4471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4588..4605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4759..5347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4839..4858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5518..6309,6310..6378))
                     /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     6141..6160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6379..6483)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6551..6569
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6607..6629)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6729..6748
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6942..6956,1..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 tcttccgacg atctccaagt
      541 cttactcaag ctcaaatcat cattcgccga ttccaatctc gccgtgttcg attcctggaa
      601 actgaactcc ggtatcggtc cttgtagctt catcggagta acctgtaatt ctagaggcaa
      661 cgttacagag attgatctct ctcgtcgagg tttatctggg aattttccat tcgattcggt
      721 ttgtgagatt cagagtctcg agaaactctc tctcggattc aattcactat ctggaataat
      781 ccccagcgat ttgaaaaact gcacgagttt gaagtatttg gatctcggca acaatttatt
      841 ctccggtgct ttccctgagt tctcatcttt aaatcagtta cagtttctgt atctcaacaa
      901 cagcgctttt tccggcgtct ttccgtggaa atcgctccga aacgcaacga gtctcgtcgt
      961 tttgagccta ggcgataatc cgtttgatgc gacggctgat ttcccggtgg aggttgtttc
     1021 tctgaagaaa ctctcgtggc tttacttgtc gaattgcagt atcgccggaa aaattccacc
     1081 ggcgattgga gatttgacgg agcttcggaa tttggagatt tccgatagcg gattaaccgg
     1141 agagattccg tcggagattt cgaaacttac taatctctgg cagttagagc tttacaacaa
     1201 ctccttgacc ggaaaacttc ctaccggatt cggaaacttg aagaatctga cttacttgga
     1261 cgcttcaaca aatcttctac aaggcgattt atcggagctt aggtctctga ctaatctcgt
     1321 ttcactacaa atgttcgaaa acgaattctc cggtgagatt ccgctggagt ttggtgaatt
     1381 caaggatctc gtgaatctct ctctgtatac aaacaagtta accggttctc tgcctcaagg
     1441 actcggttca cttgctgatt ttgatttcat cgacgcatca gagaatctct taaccggacc
     1501 gatccctcca gatatgtgca agaacgggaa gatgaaagct ctgcttctcc tgcaaaacaa
     1561 cctcactggc tctatccctg agtcctacgc taactgtttg actctgcaac ggttcagagt
     1621 tagcgagaac aacctcaacg gaactgttcc cgcgggactt tggggattac cgaagctcga
     1681 gatcatcgat atagagatga ataacttcga aggtccaatc accgcggata taaagaacgg
     1741 taaaatgctc ggagcgttgt atctagggtt caacaagttg tctgatgagc tgcctgaaga
     1801 gataggtgac actgaatctt tgactaaggt tgagcttaac aataaccggt ttaccgggaa
     1861 gattcctagc tccattggga aactcaaagg acttagcagt ctaaagatgc agagcaatgg
     1921 cttctccggt gagatacctg actcgattgg ttcgtgctcg atgctcagcg atgtcaacat
     1981 ggctcaaaac tcgatatccg gagaaatccc ccacacgctt ggatcactcc cgacgctcaa
     2041 cgctttgaat ctttcggaca ataagttgtc tgggagaatc ccagagagtt tgtcgtcttt
     2101 aaggcttagc cttcttgatc tgtctaacaa cagattatca ggtcgtatcc ctctgagttt
     2161 gtcatcatat aatggtagct tcaatggcaa tcctggcctc tgtagcacga cgatcaaatc
     2221 gtttaaccgg tgcataaatc cttcgagatc gctggaggtg ctgttccagg gaccaagcgg
     2281 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     2341 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     2401 caccgtgatg gagtgcgacg cctgcggcat gcagccctct agaggctccg gctccatcat
     2461 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     2521 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     2581 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     2641 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     2701 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     2761 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     2821 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     2881 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     2941 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     3001 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     3061 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     3121 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     3181 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     3241 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     3301 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     3361 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     3421 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     3481 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     3541 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     3601 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     3661 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     3721 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     3781 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     3841 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     3901 caccgacgcc gcgcacccgg gttctagagg aagcacctcc ggcgataacg attacaagga
     3961 tgatgatgat aaggccagcg gcaccggtca tcatcaccat caccattgag tttaaacccg
     4021 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     4081 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4141 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4201 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4261 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     4321 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4381 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4441 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4501 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4561 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4621 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4681 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5461 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5521 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5581 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5641 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5701 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5761 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5821 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5881 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5941 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6001 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6061 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6121 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6181 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6241 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6301 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6361 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6421 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6481 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6541 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     6601 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6661 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     6721 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6781 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6841 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6901 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
