LOCUS       sequence-298768-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector BRL1 B12_pECIA2 should be used with prey vector BRL1 
            B12_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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 6488)
  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: BRL1 B12_pECIA2
FEATURES             Location/Qualifiers
     source          1..6488
                     /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             2059..2070
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2741..2764
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3470..3511
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3521..3538
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3561..3578)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3611..3745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3635..3654)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3689..3708
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3889..3905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3889..3905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3902..3924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3913..3929
                     /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(3937..3943,3944..3961,3962..3967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3982..4003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4120..4137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4291..4879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4371..4390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5050..5841,5842..5910))
                     /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     5673..5692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5911..6015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6083..6101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6139..6161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6261..6280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6474..6488,1..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 gacttcaacg aaactgctct
      541 gttattggcg ttcaagcaaa actctgtaaa atctgatcct aataacgttc ttggtaactg
      601 gaaatacgag tcgggtcgtg gttcatgctc ttggcgaggg gtttcttgct ctgacgatgg
      661 tcgaatcgtc ggattagatc tccgaaacag tggactcacc ggaaccctaa acctagttaa
      721 cctcacggcg ttgcctaatc tccagaatct ttacttgcaa ggaaattact tctcctctgg
      781 aggagattcg tctggttctg attgttatct tcaagttctg gatttgtctt cgaattctat
      841 ttcagactat tcaatggtgg attacgtttt ctcaaaatgt tcgaatctgg tttcggtgaa
      901 tatctccaac aacaagctcg tcggaaaatt aggttttgct ccgtcgtctt tacagagctt
      961 gacgactgtt gatctctctt acaacatctt atcagataag attccggaga gttttatttc
     1021 agattttccg gcgtcgttga aatatctcga tctaactcac aacaacttat ccggcgactt
     1081 ctccgatctc agtttcggta tctgtgggaa tctcaccttc ttcagtctat cacagaacaa
     1141 tctctccggt gataaattcc caatcactct accaaactgc aaattcctcg agacattaaa
     1201 catctctcgg aacaatctcg ccggaaaaat ccctaacgga gagtattggg gaagtttcca
     1261 gaatctgaaa cagctctctc tagctcataa ccgtctctcc ggcgaaatcc caccggagct
     1321 ttctctactc tgcaaaaccc tagtgattct tgatctatcc ggaaacactt tctccggcga
     1381 gcttccttca cagttcaccg cctgcgtctg gttacagaat cttaacctcg gaaacaacta
     1441 cctctccgga gatttcttaa acacggtcgt gagtaaaatc accggaatca cttatctata
     1501 cgttgcttac aacaacatct caggctctgt tccgatttct ctcacaaact gttcaaatct
     1561 tcgtgttctt gatctaagct caaatggttt caccggaaat gtaccgtctg gtttctgctc
     1621 tctgcaaagc tcgccggttc ttgaaaagat tctcatagct aacaattacc tctcaggaac
     1681 agttcctatg gagcttggta aatgcaagag cttgaagaca attgatctta gcttcaatga
     1741 gcttactggt ccgattccga aagagatatg gatgttgccg aatctgtcgg atttggttat
     1801 gtgggcgaac aatctcactg ggacgatccc ggaaggtgtt tgtgttaaag gaggaaattt
     1861 ggaaactctt atcctcaaca acaatctctt aaccggttca atccctgagt cgatctcgag
     1921 atgcaccaat atgatctgga tctctctttc tagtaatcgt ctcaccggga aaatcccgag
     1981 cgggattggt aatctttcca agttagcaat cttgcagcta gggaataact ctctgtccgg
     2041 gaacgttcct cgccagcttg ggaactgcaa gagtctgatc tggcttgatc tgaacagcaa
     2101 caatctaacc ggggacctcc cgggtgagct agctagccaa gccgggttag taatgcctgg
     2161 gagcgtttca ggtaaacagt ttgcgtttgt gagaaacgaa ggtggaacag actgcagagg
     2221 tgcaggtggg ttagtagagt ttgaaggcat tcgtgctgaa cgattagagc ggctaccaat
     2281 ggttcattcg tgtcccgcga cacggatata ctcaggcatg acaatgtaca cattctcagc
     2341 aaatggaagc atgatctatt tcgacatctc gtataatgct gtttcaggtt ttatacctcc
     2401 tggttatggt aacatgggct atctccaggt cttgaatttg ggacataacc ggataaccgg
     2461 aaccatcccg gacagttttg gaggattgaa agcgattggt gttctcgatc tatctcacaa
     2521 caatcttcaa gggtacttac ccgggtcgct ggggtcgctt tctttcctca gtgatcttga
     2581 tgtctctaac aacaacctta ctggtccaat cccatttgga ggtcagctta caacgttccc
     2641 tgtctcgaga tacgcaaaca actcaggcct ctgtggtgtt cctttgcgtc cctgcggttc
     2701 agctcctcga cggcccatta cctcccgtat ccatgccaag ctggaggtgc tgttccaggg
     2761 accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     2821 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     2881 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     2941 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     3001 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     3061 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     3121 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     3181 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     3241 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     3301 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     3361 gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca accactacac
     3421 gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag gtaagcctat
     3481 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     3541 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     3601 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3661 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3721 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3781 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3841 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3901 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3961 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4021 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4081 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4141 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4201 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4261 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4321 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4381 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4441 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4501 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4561 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4621 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4681 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4741 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4801 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4861 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4921 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5041 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5101 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5161 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5221 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5281 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5341 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5401 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5461 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5521 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5581 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5641 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5701 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5761 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5821 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5881 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5941 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6001 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6061 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6121 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6181 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6241 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6301 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6481 acgacgtt
//
