LOCUS       sequence-298776-        6506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector BRL3 C7_pECIA2 should be used with prey vector BRL3 
            C7_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6506)
  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 6506)
  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: BRL3 C7_pECIA2
FEATURES             Location/Qualifiers
     source          1..6506
                     /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             2759..2782
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3488..3529
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3539..3556
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3579..3596)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3629..3763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3653..3672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3707..3726
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3907..3923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3907..3923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3920..3942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3931..3947
                     /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(3955..3961,3962..3979,3980..3985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4000..4021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4138..4155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4309..4897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4389..4408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5068..5859,5860..5928))
                     /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     5691..5710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5929..6033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6101..6119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6157..6179)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6279..6298
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6492..6506,1..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 agacggctac tcagcgacga
      541 cgttaacgac acagctctgc taacggcgtt taagcaaacc tctatcaagt cggatcctac
      601 caacttttta ggtaactgga gatacgggtc gggtcgtgac ccatgtactt ggcgaggcgt
      661 ctcttgttct agcgatggta gagtcattgg acttgatcta cgaaacggtg gtcttaccgg
      721 aaccctaaat ctcaataatc tcacggcgtt gtctaatctc cggagtcttt atttgcaagg
      781 aaacaacttc tcctccggag attcttcttc ttcttcagga tgttctctcg aggttctgga
      841 tttatcttca aactcgctta cggactcttc aatagtggat tacgttttct ccacctgttt
      901 aaatctggtt tctgtcaatt tctcgcacaa caagctcgcc gggaaattga agtcttctcc
      961 ttcggcgagt aacaagagaa tcacaaccgt tgatctctcg aacaatcgct tctccgatga
     1021 gattccggag actttcatcg ccgattttcc gaattctctg aaacacctcg acttatccgg
     1081 caacaatgtc accggcgact tctctcgtct cagctttggt ctctgtgaga acctcaccgt
     1141 attcagctta tcacagaaca gcatctccgg cgacagattt ccggttagtc tctcaaattg
     1201 taagctcctc gaaacgttaa acctctctcg gaacagtctc atcggaaaaa tccccggaga
     1261 tgactattgg ggaaattttc agaacctgag gcagctctct cttgcacaca acctttactc
     1321 cggtgagatt ccaccggagc tttctctgct ctgtcgaact ctagaggttc tcgatctctc
     1381 cggcaacagt ctcaccggtc agcttccgca atcattcacc tcttgtggct cattacaaag
     1441 cctaaatctt ggaaacaaca agctctccgg tgatttctta agcaccgtcg tgagtaaact
     1501 ctctagaatc actaatcttt acttaccatt caacaacatc tcaggctctg ttccgatttc
     1561 gctcaccaac tgtagtaatc tgcgagttct tgatctgagc tcaaacgagt tcactggaga
     1621 agtaccatct ggtttctgct ctcttcaaag ctcttcggtt ttggaaaagt tgcttattgc
     1681 caacaattac ctctcaggaa ctgtccctgt agagcttggt aaatgtaaga gcttgaagac
     1741 tatagatctt agcttcaatg ctctcactgg tctgattcct aaggagatat ggacattgcc
     1801 taagctctcc gacttagtta tgtgggcgaa caatctcact ggtggaatcc cggagagcat
     1861 ttgtgttgat ggaggaaact tggaaacgtt aatcttaaac aacaatctct taaccggttc
     1921 tcttccagaa tcaatctcca aatgcaccaa catgctttgg atctcccttt ctagtaacct
     1981 gcttaccggt gaaatccctg taggaatcgg aaagcttgag aagttagcga ttctgcaact
     2041 cgggaacaat tctttaactg ggaacattcc aagtgagctt ggaaactgca agaaccttat
     2101 ctggcttgat cttaacagca ataatctaac tgggaatctc ccaggtgaac ttgcaagtca
     2161 ggccggactg gtaatgcccg gaagcgtctc tggtaaacag tttgcgtttg tgaggaatga
     2221 aggtgggacg gattgcagag gtgcaggggg tttggttgag tttgaaggaa ttcgtgctga
     2281 gcggttagag cattttccaa tggttcattc atgtcccaag actcggatat actcaggcat
     2341 gaccatgtac atgttctcca gtaacgggag catgatctac ttggaccttt cctacaatgc
     2401 ggtttcaggt tctattcctt tgggttatgg tgcaatggga tatcttcagg tcttgaattt
     2461 gggacataac ttgttaactg gaaccatacc ggatagcttc ggaggattga aagcaattgg
     2521 agttcttgat ctatctcaca atgatctcca aggattctta cctggatcac taggaggact
     2581 ctcttttcta agcgacttag acgtctctaa caacaatcta accggaccaa tcccattcgg
     2641 aggccaactt acgacgttcc cactcacaag atacgccaac aactctggcc tctgcggggt
     2701 tcctctacca ccttgtagct ctggttcccg gcccacgaga tctcatgctc accctaaact
     2761 ggaggtgctg ttccagggac cagggagcga caaaactcac acatgcccac cgtgcccagc
     2821 acctgaactc ctggggggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct
     2881 catgatctcc cggacccctg aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc
     2941 tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc
     3001 gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca
     3061 ggactggctg aatggcaagg agtacaagtg caaggtctcc aacaaagccc tcccagcccc
     3121 catcgagaaa accatctcca aagccaaagg gcagccccga gaaccacagg tgtacaccct
     3181 gcccccatcc cgggaggaga tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg
     3241 cttctatccc agcgacatcg ccgtggagtg ggagagcaat gggcagccgg agaacaacta
     3301 caagaccacg cctcccgtgc tggactccga cggctccttc ttcctctaca gcaagctcac
     3361 cgtggacaag agcaggtggc agcaggggaa cgtcttctca tgctccgtga tgcatgaggg
     3421 tctgcacaac cactacacgc agaagagcct ctccctgtct ccgggtaaat ctagagggcc
     3481 cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     3541 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     3601 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3661 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3721 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3781 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3841 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     3901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6061 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6121 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6181 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6241 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6301 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6361 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6421 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6481 gccagggttt tcccagtcac gacgtt
//
