LOCUS       sequence-298772-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector BRL2 X039_pECIA2 should be used with prey vector BRL2 
            X039_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  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 6404)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6404
                     /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             3386..3427
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3437..3454
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3477..3494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3527..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3551..3570)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3605..3624
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3805..3821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3805..3821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3818..3840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3829..3845
                     /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(3853..3859,3860..3877,3878..3883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3898..3919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4036..4053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4207..4795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4287..4306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4966..5757,5758..5826))
                     /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     5589..5608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5827..5931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5999..6017
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6055..6077)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6177..6196
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6390..6404,1..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 gggagcgaca aaactcacac
     2701 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc
     2761 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga
     2821 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca
     2881 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt
     2941 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa
     3001 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga
     3061 accacaggtg tacaccctgc ccccatcccg ggaggagatg accaagaacc aggtcagcct
     3121 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg
     3181 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt
     3241 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg
     3301 ctccgtgatg catgagggtc tgcacaacca ctacacgcag aagagcctct ccctgtctcc
     3361 gggtaaatct agagggccct tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     3421 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     3481 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     3541 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3601 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3661 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3721 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgacctgcag
     3781 gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5941 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6001 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6061 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6121 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6181 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6241 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     6301 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6361 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
