LOCUS       sequence-274520-        6968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RKF1 X030_pECIA14 should be used with bait vector RKF1 
            X030_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6968)
  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 6968)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..6968
                     /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             3962..3985
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4001..4018
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4041..4058)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4091..4225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4115..4134)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4169..4188
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4369..4385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4369..4385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4382..4404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4393..4409
                     /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(4417..4423,4424..4441,4442..4447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4462..4483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4600..4617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4771..5359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4851..4870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5530..6321,6322..6390))
                     /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     6153..6172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6391..6495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6563..6581
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6619..6641)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6741..6760
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6954..6968,1..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 gggaccaagc ggcaatcccg gcggcgatct ggccccccag atgctgcgcg agctgcagga
     2341 gaccaatgcc gccctgcagg acgtgcgcga gctgctgcgc caccgcgtga aggagatcac
     2401 cttcctgaag aacaccgtga tggagtgcga cgcctgcggc atgcagccct ctagaggctc
     2461 cggctccatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     2521 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     2581 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     2641 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     2701 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     2761 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     2821 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     2881 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     2941 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     3001 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     3061 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     3121 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     3181 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     3241 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     3301 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     3361 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     3421 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     3481 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     3541 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     3601 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     3661 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     3721 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     3781 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     3841 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     3901 ccccgccggc accaccgacg ccgcgcaccc gggttctaga ggaagcacct ccggcgataa
     3961 cgattacaag gatgatgatg ataaggccag cggcaccggt catcatcacc atcaccattg
     4021 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     4081 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4141 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4201 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4261 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     4321 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     4381 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4441 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4501 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4561 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4621 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4681 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4741 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4801 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4861 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4921 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4981 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5041 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5101 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5161 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5221 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5281 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5341 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5401 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5461 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5521 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5581 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5641 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5701 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5761 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5821 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5881 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5941 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6001 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6061 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6121 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6181 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6241 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6301 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6361 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6421 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6481 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6541 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6601 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6661 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6721 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6781 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6841 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6901 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6961 acgacgtt
//
