LOCUS       sequence-275921-        7361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector BRL2 X039_pECIA14 should be used with bait vector BRL2 
            X039_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7361)
  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 7361)
  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: BRL2 X039_pECIA14
FEATURES             Location/Qualifiers
     source          1..7361
                     /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             2657..2680
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4355..4378
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4394..4411
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4434..4451)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4484..4618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4508..4527)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4562..4581
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4762..4778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4762..4778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4775..4797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4786..4802
                     /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(4810..4816,4817..4834,4835..4840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4855..4876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4993..5010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5164..5752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5244..5263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5923..6714,6715..6783))
                     /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     6546..6565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6784..6888)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6956..6974
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7012..7034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7134..7153
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7347..7361,1..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 tcttcactca aaaccgactc
      541 attatcactc ctctctttca aaacaatgat ccaagatgat ccaaacaaca ttctctcaaa
      601 ctggtcacct cgtaaatccc cttgtcaatt ctccggcgtc acttgtctcg gaggaagagt
      661 caccgagatt aatctctccg gtagtggcct ctccggaatc gtctccttca atgctttcac
      721 ctctcttgat tctctctctg ttctcaaact ctcagagaac ttcttcgtct taaactcaac
      781 ctctcttctt cttcttcctt taaccctaac tcatctcgaa ctctcttcct ccggtttaat
      841 cggaaccctc cctgaaaatt tcttctccaa atactcaaat cttatttcca tcaccctctc
      901 ttacaataac ttcaccggaa agttacctaa tgatcttttc ttaagcagca agaaactcca
      961 aacccttgat ctttcttaca acaacattac cggtccaatc tccggtttaa ccattcctct
     1021 ttcttcttgc gtctccatga cctatctcga tttctccggt aacagtatct ccggctatat
     1081 ctctgactca ctcatcaact gcaccaacct caaaagcttg aatctttctt acaacaactt
     1141 cgatgggcag attccaaaat ctttcggtga gcttaaactc ttacaaagtt tggatctttc
     1201 tcataaccga ctcaccggtt ggatcccgcc ggagatcgga gatacttgcc ggtcattaca
     1261 aaacctccgg ctttcttaca acaacttcac tggtgtgatc cctgagtcgt tgtcctcatg
     1321 ttcttggcta caaagtcttg acctttcaaa caacaacatc tccggtccgt ttccgaacac
     1381 tatcctccgg agttttggat ctcttcaaat cttacttctc agcaacaact taatctccgg
     1441 tgactttccg acttcaatct ctgcttgtaa gagcttgaga atcgctgact tcagctcgaa
     1501 tcgattctcc ggtgtgattc caccggattt atgtcccgga gcagcttcat tagaagagct
     1561 aagactcccg gataatctcg tcaccggaga gattccaccg gcgatatctc aatgctctga
     1621 gctacgaacg attgatctca gtctcaatta tctcaacggt acaatcccac cggagatcgg
     1681 aaacctccag aaactcgagc aattcatagc ttggtacaac aacatcgccg gagaaatccc
     1741 accggagatc ggaaaactac aaaacctaaa agatttgatc ctcaacaaca atcaactcac
     1801 cggagaaatc ccaccggaat ttttcaattg cagtaacatc gaatgggtct cattcacaag
     1861 caacagatta accggagaag tgcctaaaga tttcggaatc ctttcccggt tagccgttct
     1921 tcagctcggt aacaacaatt tcaccggaga aatcccaccg gagctaggaa aatgcacaac
     1981 tctggtttgg cttgatctta acacaaacca cttaaccggt gaaatccctc cccggttagg
     2041 tcgtcaaccc ggttcaaaag cactttccgg tttactctca ggaaacacaa tggcttttgt
     2101 tagaaacgtt ggaaactcat gtaaaggtgt tggaggtttg gttgagttct cagggatcag
     2161 accagagagg cttcttcaga ttccatctct taaaagctgt gacttcacaa gaatgtattc
     2221 aggtccaatt ttgagtctat tcacaagata tcaaaccatt gagtatcttg atctttctta
     2281 taatcagctt agaggtaaaa tccctgatga gattggagaa atgattgctc ttcaagttct
     2341 tgagctttct cataatcaac tctccggcga gattccgttt acgattggtc agctcaagaa
     2401 tctcggtgtt ttcgatgctt cggataatcg gttgcagggt caaatccctg aatctttctc
     2461 gaatttgtct ttcttggtgc agattgattt gtcgaataac gagttaactg gtccgattcc
     2521 tcagagaggt cagttaagca cgcttccggc gactcagtac gcgaataatc cggggctttg
     2581 cggtgttcct ttaccggaat gtaagaatgg aaacaatcag cttcctgctg gaacagagga
     2641 agggaaacgt gctaagctgg aggtgctgtt ccagggacca agcggcaatc ccggcggcga
     2701 tctggccccc cagatgctgc gcgagctgca ggagaccaat gccgccctgc aggacgtgcg
     2761 cgagctgctg cgccaccgcg tgaaggagat caccttcctg aagaacaccg tgatggagtg
     2821 cgacgcctgc ggcatgcagc cctctagagg ctccggctcc atcatcccag ttgaggagga
     2881 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     2941 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     3001 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     3061 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     3121 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     3181 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     3241 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     3301 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     3361 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     3421 tacgcagctc atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat
     3481 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     3541 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     3601 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     3661 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     3721 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     3781 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     3841 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     3901 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     3961 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     4021 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     4081 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     4141 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     4201 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     4261 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     4321 cccgggttct agaggaagca cctccggcga taacgattac aaggatgatg atgataaggc
     4381 cagcggcacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     4441 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     4501 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4561 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4621 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4681 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga cctgcaggca
     4741 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4801 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4861 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4921 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4981 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5041 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5101 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5161 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5281 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5341 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5401 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5461 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5521 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5581 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5641 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5701 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5761 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5821 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5881 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5941 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6001 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6061 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6121 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6181 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6241 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6301 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6361 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6421 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6481 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6541 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6601 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6661 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6721 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6781 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6841 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6901 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6961 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7021 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7081 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7141 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7201 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7261 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     7321 attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
