LOCUS       sequence-298800-        5999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RLK7/KUK2 C12_pECIA2 should be used with prey vector 
            RLK7/KUK2 C12_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5999)
  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 5999)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..5999
                     /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             2981..3022
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3032..3049
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3072..3089)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3122..3256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3146..3165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3200..3219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3400..3416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3400..3416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3413..3435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3424..3440
                     /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(3448..3454,3455..3472,3473..3478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3493..3514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3631..3648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3802..4390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3882..3901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4561..5352,5353..5421))
                     /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     5184..5203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5422..5526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5594..5612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5650..5672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5772..5791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5985..5999,1..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 gaccagggag
     2281 cgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg gaccgtcagt
     2341 cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac
     2401 atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga
     2461 cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca acagcacgta
     2521 ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca aggagtacaa
     2581 gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct ccaaagccaa
     2641 agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg agatgaccaa
     2701 gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca tcgccgtgga
     2761 gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc
     2821 cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt ggcagcaggg
     2881 gaacgtcttc tcatgctccg tgatgcatga gggtctgcac aaccactaca cgcagaagag
     2941 cctctccctg tctccgggta aatctagagg gcccttcgaa ggtaagccta tccctaaccc
     3001 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     3061 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     3121 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3181 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3241 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3301 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     3361 tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3421 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5581 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5641 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5701 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5761 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5821 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     5881 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     5941 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
