LOCUS       sequence-274762-        7445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector BRL1 B12_pECIA14 should be used with bait vector BRL1 
            B12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7445)
  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 7445)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..7445
                     /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             4439..4462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4478..4495
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4518..4535)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4568..4702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4592..4611)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4646..4665
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4846..4862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4846..4862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4859..4881)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4870..4886
                     /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(4894..4900,4901..4918,4919..4924))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4939..4960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5077..5094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5248..5836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5328..5347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6007..6798,6799..6867))
                     /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     6630..6649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6868..6972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7040..7058
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7096..7118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7218..7237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7431..7445,1..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 accaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc tgcaggagac
     2821 caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt
     2881 cctgaagaac accgtgatgg agtgcgacgc ctgcggcatg cagccctcta gaggctccgg
     2941 ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg cagccgaggc
     3001 cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc tcatcatctt
     3061 cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa aagggcagaa
     3121 gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat atgtggctct
     3181 gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag ccacggccta
     3241 cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg cccgctttaa
     3301 ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg ccaagaaagc
     3361 agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc cagccggcac
     3421 ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg cctcggcccg
     3481 ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca ttgacgtgat
     3541 cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg agtacccaga
     3601 tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg aatggctggc
     3661 gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg cttccctgga
     3721 cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat acgagatcca
     3781 ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc tgcgcctgct
     3841 gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg accatggtca
     3901 tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg acgccattga
     3961 gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg ccgaccactc
     4021 ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg ggctggcccc
     4081 tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg gtccaggcta
     4141 tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga gccccgagta
     4201 tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg acgtggcggt
     4261 gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga ccttcatagc
     4321 gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc tggcgccccc
     4381 cgccggcacc accgacgccg cgcacccggg ttctagagga agcacctccg gcgataacga
     4441 ttacaaggat gatgatgata aggccagcgg caccggtcat catcaccatc accattgagt
     4501 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     4561 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4621 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4681 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4741 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     4801 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     4861 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4921 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4981 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5041 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5101 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5161 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5221 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5281 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5341 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5401 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5461 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5521 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5581 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5641 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5701 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5761 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5821 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5881 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5941 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6001 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6061 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6121 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6181 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6241 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6301 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6361 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6421 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6481 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6541 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6601 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6661 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6721 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6781 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6841 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6901 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6961 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7021 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     7081 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7141 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7201 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     7261 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     7321 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     7381 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     7441 acgtt
//
