LOCUS       sequence-298777-        6617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector BRI1 BRI1_pECIA2 should be used with prey vector BRI1 
            BRI1_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6617)
  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 6617)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6617
                     /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             3599..3640
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3650..3667
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3690..3707)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3740..3874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3764..3783)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3818..3837
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4031..4053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4042..4058
                     /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(4066..4072,4073..4090,4091..4096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4111..4132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4249..4266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4420..5008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4500..4519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5179..5970,5971..6039))
                     /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     5802..5821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6040..6144)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6212..6230
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6268..6290)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6390..6409
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6603..6617,1..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 ccagggagcg acaaaactca cacatgccca ccgtgcccag cacctgaact
     2941 cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc
     3001 ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa
     3061 gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga
     3121 gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct
     3181 gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa
     3241 aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc
     3301 ccgggaggag atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc
     3361 cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac
     3421 gcctcccgtg ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa
     3481 gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcatgagg gtctgcacaa
     3541 ccactacacg cagaagagcc tctccctgtc tccgggtaaa tctagagggc ccttcgaagg
     3601 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     3661 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     3721 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     3781 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3841 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3901 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3961 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggta
     4021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6181 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6241 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6301 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6361 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6421 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6481 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6541 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6601 ttcccagtca cgacgtt
//
