LOCUS       sequence-238694-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector FEI2 X041_pECIA14 should be used with bait vector FEI2 
            X041_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  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 5855)
  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: FEI2 X041_pECIA14
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1596..1619
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3294..3317
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3333..3350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3373..3390)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3423..3557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3447..3466)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3501..3520
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3701..3717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3701..3717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3714..3736)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3725..3741
                     /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(3749..3755,3756..3773,3774..3779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3794..3815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3932..3949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4103..4691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4183..4202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4862..5653,5654..5722))
                     /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     5485..5504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5723..5827)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggaatga agcaattagt cctgatggtg aggcgctttt gagttttaga aatggagttt
     1021 tggcttctga tggtgttatt ggtctgtgga gaccagagga tcctgatcca tgtaactgga
     1081 aaggagtgac ctgtgatgca aaaacaaaaa gagttatagc cttgagtctt acttatcaca
     1141 aattaagggg acctttaccc cctgagcttg gaaagctgga tcagttaagg cttttaatgc
     1201 ttcacaacaa tgcgttatat caatcaatac ctgcctcatt gggaaattgc acggcattag
     1261 agggaatata cttgcagaat aattacatca ctgggacaat cccaagtgaa ataggaaacc
     1321 tgtccgggct gaaaaacctg gacctctcaa acaacaatct caatggagct atccctgctt
     1381 cacttgggca gttgaaaagg cttactaaat tcaatgtctc aaacaatttc ctggtgggaa
     1441 aaataccttc tgacggccta ctcgctagac tctcgaggga ctccttcaac ggaaatcgta
     1501 acttgtgtgg aaaacaaatc gatattgtgt gcaatgacag tggaaattct acagcttctg
     1561 ggtctccaac aggccaagga ggtaataacc ctaaactgga ggtgctgttc cagggaccaa
     1621 gcggcaatcc cggcggcgat ctggcccccc agatgctgcg cgagctgcag gagaccaatg
     1681 ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt gaaggagatc accttcctga
     1741 agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc ctctagaggc tccggctcca
     1801 tcatcccagt tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg
     1861 gtgccgccaa gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg
     1921 gcgatgggat gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg
     1981 acaaactggg gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca
     2041 agacatacaa tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt
     2101 gcggggtcaa gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt
     2161 gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga
     2221 agtcagtggg agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg
     2281 cccacacggt gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg
     2341 aggggtgcca ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag
     2401 gtggaggccg aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact
     2461 acagccaagg tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc
     2521 gccagggtgc ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt
     2581 ctgtgaccca tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag
     2641 actccacact ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca
     2701 ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg
     2761 aaagcagggc ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg
     2821 cgggccagct caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg
     2881 tcttctcctt cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca
     2941 aggcccggga caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc
     3001 tcaaggacgg cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc
     3061 agcagtcagc agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg
     3121 cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg
     3181 tcatggcctt cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg
     3241 gcaccaccga cgccgcgcac ccgggttcta gaggaagcac ctccggcgat aacgattaca
     3301 aggatgatga tgataaggcc agcggcaccg gtcatcatca ccatcaccat tgagtttaaa
     3361 cccgctgatc agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg
     3421 aggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3481 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3541 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3601 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     3661 atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3721 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3781 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3841 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3901 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4861 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4921 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4981 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5041 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5101 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5161 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5221 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5281 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5341 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5401 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5461 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5521 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5581 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5641 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5701 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5761 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5821 ttccgcgcac atttccccga aaagtgccac ctgac
//
