LOCUS       sequence-297035-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RKF1 X030_pECIA2 should be used with prey vector RKF1 
            X030_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  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 6011)
  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: RKF1 X030_pECIA2
FEATURES             Location/Qualifiers
     source          1..6011
                     /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             2264..2287
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2993..3034
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3044..3061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3084..3101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3134..3268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3158..3177)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3212..3231
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3412..3428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3412..3428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3425..3447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3436..3452
                     /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(3460..3466,3467..3484,3485..3490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3505..3526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3643..3660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3814..4402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3894..3913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4573..5364,5365..5433))
                     /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     5196..5215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5434..5538)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5606..5624
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5662..5684)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5784..5803
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5997..6011,1..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 aaattaccgc agcaagaagt
      541 cgatgctctg cagcaaatcg ctaccacatt gggctcgaaa ttctggaaat tcgatgctga
      601 aaattgtaag attgaaatgg tggggttaac tgaaactcct cctccaacag ctaagcaaga
      661 aatagaatgt gaatgcagtc ccactaacga cactgattgt catgttgtca aatttgcatt
      721 caaggaccat aatcttccag gaactcttcc tcaaatagtc aagcttccct accttcggga
      781 gatagacttg gcctacaact acatcaatgg gaccttacct cgtgaatggg cttcttcaaa
      841 tttaactttt atttctcttc ttgtgaatcg gttgtctgga gaaattccga aggagtttgg
      901 aaactcttct ctaacatact tggatctcga gtcaaatgcg ttttctggaa ctattcctca
      961 ggagcttggg aacttagtcc acttgaaaaa actgttgctt tcctctaaca aattgaccgg
     1021 aactttgcca gcttcactag ctagactaca gaatatgaca gatttcagga ttaacgatct
     1081 tcaactcagt ggaacgatac ctagttacat acaaaattgg aagcaactcg agagacttga
     1141 aatgattgca tctggtctca ctggtccaat cccatcggtc atctcagttc tgagcaatct
     1201 tgttaatctg aggatcagtg atatacgtgg accagtccaa ccattccctt cgttgaagaa
     1261 cgtgacaggt ctaactaaaa ttatattgaa gaactgtaac atttctggac aaattcctac
     1321 ttatctttca catttgaagg agttggagac actggatttg agttttaaca agttggttgg
     1381 gggaattccg tcgtttgcac aagcagaaaa tttacggttt ataattttgg ctggaaatat
     1441 gctcgaggga gatgctccag atgaattact aagagatgga atcacagtag atctttcata
     1501 taacaatctc aagtggcaga gtcctgaaag tcgtgcttgt cgcccaaata tgaatctgaa
     1561 tctaaacttg tttcagagca cttcgacaaa aaaatcgagc aaatttctgc cgtgtatcaa
     1621 agatttcaaa tgtcctcgat attctagctg cttgcatgtg aactgcggtg gaagcgatat
     1681 gtatgttaag gagaaaaaaa caaaagaatt atatgaagga gatggaaatg ttgaaggtgg
     1741 tgctgctaag tattttttga agcctgacgc taattgggga tttagcagca caggggactt
     1801 catggacgat aacaatttcc aaaacactag attcaccatg tttgtcccgg catctaacca
     1861 gtctgatctc tacaaatcgg cacgaattgc tccagtatct ctaacttact tccatgcctg
     1921 cttggaaaat ggaaattaca ctataaatct agacttcgct gagatacggt tcaccaatga
     1981 tgaaaactat aaccggcttg gaaggcgtct gtttgacatt tacattcagg agaaattagt
     2041 ggcaaaggac tttaacatca tggatgaagc aaagggagct caaacaccta taatcaaacc
     2101 acttactgcc tatgtgacaa accacttttt aaccatccgg ttgagttggg ccggtaaagg
     2161 aactaccagg attcccacta gaggggttta tggtcctatc atatcggcta tttccatagt
     2221 ttcagattcc aagccgtgcg aacgtccgaa aactggaatg agcctggagg tgctgttcca
     2281 gggaccaggg agcgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg
     2341 gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac
     2401 ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa
     2461 ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta
     2521 caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg
     2581 caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
     2641 ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga
     2701 ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga
     2761 catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc
     2821 cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag
     2881 gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gagggtctgc acaaccacta
     2941 cacgcagaag agcctctccc tgtctccggg taaatctaga gggcccttcg aaggtaagcc
     3001 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     3061 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     3121 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     3181 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3241 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3301 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3361 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     3421 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3481 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3541 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3601 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4561 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4741 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5101 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5401 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5581 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5641 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5701 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5761 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5821 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     5881 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5941 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6001 gtcacgacgt t
//
