LOCUS       sequence-275965-        7574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector BRI1 BRI1_pECIA14 should be used with bait vector BRI1 
            BRI1_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7574)
  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 7574)
  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: BRI1 BRI1_pECIA14
FEATURES             Location/Qualifiers
     source          1..7574
                     /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"
     sig_peptide     521..580
                     /codon_start=1
                     /gene="Autographa californica gp64"
                     /product="signal sequence from the baculovirus envelope 
                     glycoprotein GP64"
                     /label=GP64 signal sequence
                     /translation="MVSAIVLYVLLAAAAHSAFA"
     CDS             complement(2447..2458)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2870..2893
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4568..4591
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4607..4624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4647..4664)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4697..4831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4721..4740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4775..4794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4975..4991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4975..4991)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4988..5010)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4999..5015
                     /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(5023..5029,5030..5047,5048..5053))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5068..5089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5206..5223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5377..5965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5457..5476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6136..6927,6928..6996))
                     /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     6759..6778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6997..7101)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7169..7187
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7225..7247)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7347..7366
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7560..7574,1..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 atggtaagcg ctattgtttt
      541 atatgtgctt ttggcggcgg cggcgcattc tgcctttgcg gccatgggta gccagagcct
      601 gtaccgcgaa attcaccagc tgatttcttt caaagatgtg ctgccggaca aaaacctgct
      661 gccggattgg agcagcaaca aaaacccgtg caccttcgat ggcgtgacct gccgtgatga
      721 taaagtgacc agcatcgatc tgagcagcaa accgctgaac gtgggcttca gcgccgtgag
      781 cagcagcctg ctgtctctga ccggcctgga aagcctgttc ctgagcaaca gccacatcaa
      841 cggcagcgtg agcggcttca aatgcagcgc cagcctgacc agcctggatc tgtctcgcaa
      901 cagcctgagc ggtccggtga ccaccctgac cagtctgggc agctgcagcg gcctgaaatt
      961 cctgaacgtg tccagcaaca ccctggattt cccgggcaaa gtgtctggcg gtctgaaact
     1021 gaacagcctg gaagtgctgg atctgagcgc caacagcatc agcggcgcca acgtggtggg
     1081 ctgggtgctg tctgatggct gcggcgaact gaaacacctg gccatcagcg gcaacaaaat
     1141 cagcggtgat gtggatgtga gccgctgcgt gaacctggaa ttcctggatg tgtcctccaa
     1201 caacttcagc accggcatcc cgtttctggg cgattgcagc gccctgcagc acctggatat
     1261 ctctggcaac aaactgagcg gcgatttcag ccgcgccatc agcacctgca cggaactgaa
     1321 actgctgaac atcagcagca accagttcgt gggcccaatt ccgccactgc cgctgaaaag
     1381 cctgcagtac ctgagcctgg ccgaaaacaa attcaccggc gaaatcccgg atttcctgag
     1441 cggcgcctgc gataccctga cgggcctgga cctgagcggc aaccacttct acggcgccgt
     1501 gccgccgttc ttcggtagct gctctctgct ggaaagtctg gccctgtcta gcaacaactt
     1561 ctctggcgaa ctgccgatgg ataccctgct gaaaatgcgt ggtctgaaag tcctggacct
     1621 gtccttcaac gaattcagcg gtgagctgcc ggaatctctg accaacctga gcgcctctct
     1681 gctgaccctg gatctgagtt cgaacaactt cagtggcccg atcctgccga acctgtgcca
     1741 gaacccgaaa aacaccctgc aagaactgta cctgcagaac aacggcttca ccggcaaaat
     1801 tccgccgacc ctgagcaact gcagcgaact ggtgtctctg cacctgagct tcaactacct
     1861 gtctggcacc atcccgagca gcctgggcag cctgagcaaa ctgcgtgatc tgaaactgtg
     1921 gctgaacatg ctggaaggcg aaattccgca agaactgatg tacgtcaaaa ccctggaaac
     1981 cctgattctg gacttcaacg atctgaccgg tgaaattccg agcggcctgt ctaactgcac
     2041 caacctgaac tggatcagcc tgagtaacaa ccgcctgacg ggtgaaatcc cgaaatggat
     2101 cggccgcctg gaaaacctgg cgatcctgaa actgtccaac aacagcttca gtggtaatat
     2161 tccagcggaa ctgggcgact gtcgtagcct gatctggctg gacctgaaca ccaacctgtt
     2221 caacggcacc attccggccg ctatgttcaa acagagcggc aaaatcgccg ccaacttcat
     2281 tgccggcaaa cgctacgtgt acatcaaaaa cgatggcatg aaaaaagaat gccacggtgc
     2341 cggcaacctg ctggaatttc agggcatccg cagcgaacag ctgaaccgcc tgagcacccg
     2401 caacccgtgc aacatcacca gccgcgtgta cggcggccac accagcccga ccttcgataa
     2461 caacggctct atgatgttcc tggatatgag ctacaacatg ctgtctggct acattccgaa
     2521 agaaatcggc agcatgccgt acctgttcat cctgaacctg ggccacaacg atatcagcgg
     2581 cagcatcccg gatgaagtgg gcgatctgcg cggtctgaac attctggatc tgtcgagcaa
     2641 caaactggat ggccgcatcc cgcaggccat gagcgccctg accatgctga ccgaaatcga
     2701 cctgtcgaac aacaacctgt ccggcccgat tccggaaatg ggccagttcg aaacctttcc
     2761 gccggctaaa tttctgaaca acccgggcct gtgcggctac ccgctgccgc gctgcgatcc
     2821 gagcaacgcc gatggctacg cccaccatca gcgcagccac ggccgtcgcc tggaggtgct
     2881 gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc tgcgcgagct
     2941 gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc gcgtgaagga
     3001 gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc agccctctag
     3061 aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga accgcgaggc
     3121 agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg ccaagaacct
     3181 catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca ggatcctaaa
     3241 agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc gcttcccata
     3301 tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg gagccacagc
     3361 cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga gtgcagccgc
     3421 ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga tgaatcgggc
     3481 caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc acgcctcgcc
     3541 agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg acgtgcctgc
     3601 ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca acatggacat
     3661 tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc cagaccctga
     3721 gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc tggtgcagga
     3781 atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc tcatgcaggc
     3841 ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag acatgaaata
     3901 cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag aggctgccct
     3961 gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg gtcgcatcga
     4021 ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca tgttcgacga
     4081 cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc tcgtcactgc
     4141 cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct ccatcttcgg
     4201 gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat acggaaacgg
     4261 tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg agagcgggag
     4321 ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg caggcgagga
     4381 cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc aggagcagac
     4441 cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg cctgcgacct
     4501 ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa gcacctccgg
     4561 cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc atcaccatca
     4621 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     4681 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     4741 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4801 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4861 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4921 tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     4981 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5161 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5881 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5941 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6001 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6061 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6121 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6181 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6241 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6301 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6361 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6421 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6481 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6541 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6601 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6661 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6721 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6781 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6841 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6901 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6961 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7021 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7081 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7141 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     7201 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     7261 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     7321 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     7381 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     7441 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7501 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7561 ccagtcacga cgtt
//
