LOCUS       sequence-298860-        6116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector EFR D5_pECIA2 should be used with prey vector EFR 
            D5_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  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 6116)
  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: EFR D5_pECIA2
FEATURES             Location/Qualifiers
     source          1..6116
                     /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             2369..2392
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3098..3139
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3149..3166
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3189..3206)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3239..3373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3263..3282)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3317..3336
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3517..3533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3517..3533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3530..3552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3541..3557
                     /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(3565..3571,3572..3589,3590..3595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3610..3631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3748..3765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3919..4507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3999..4018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4678..5469,5470..5538))
                     /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     5301..5320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5539..5643)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5711..5729
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5767..5789)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5889..5908
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6102..6116,1..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 tctaatgaga ctgatatgca
      541 agctttgctt gagttcaagt ctcaagtttc tgaaaacaac aagagagagg tcttggcttc
      601 atggaatcac tcctccccat tttgtaattg gattggagtc acatgtggcc gcaggagaga
      661 aagagttata agtttgaacc ttggaggatt caagttaacc ggtgtgatct caccctccat
      721 tggtaatctc tcctttctta gattacttaa tcttgcagac aactcttttg gaagtaccat
      781 ccctcaaaag gtgggaaggc tatttaggct tcagtacttg aacatgagct ataatcttct
      841 cgaaggaagg attccgtcta gtctttctaa ctgctctaga ctgtcgaccg ttgatttatc
      901 gtcaaaccat cttggacatg gtgttccttc agaactaggt tcactttcta agcttgccat
      961 tctggatctt agcaaaaaca accttactgg aaattttcct gcatctttag gaaacttgac
     1021 gtcacttcag aagcttgact ttgcatataa ccagatgaga ggtgagattc cagacgaagt
     1081 agctagattg actcaaatgg tgtttttcca aatagcactg aatagttttt caggtggttt
     1141 tcctcctgca ttgtacaaca tctcctctct tgagtctcta tctctagctg acaatagctt
     1201 ttcgggtaat cttagggctg attttggtta tcttctacca aatctaagaa gacttctttt
     1261 gggaacaaat cagttcactg gagctattcc caaaacactt gccaatatct caagccttga
     1321 aaggtttgat atctcatcta attacctgtc tggtagtatc cctttgagct ttggaaagtt
     1381 acgtaatctg tggtggttag ggattcgtaa taactctctt ggaaataact cgtccagtgg
     1441 tcttgaattt attggggctg tggcgaactg cactcaatta gagtacttag atgttggtta
     1501 caatagactt ggaggtgagc ttcctgcttc tatagccaat ctgtccacta cattgactag
     1561 tctgttcctt ggacaaaatc ttatctctgg aaccattcct catgacatcg ggaatcttgt
     1621 aagcctgcaa gaactcagct tagaaacaaa tatgttgagt ggagaacttc ccgtctcttt
     1681 cgggaagctt ttgaacttgc aggttgtgga tctgtattca aatgcaatat cgggggaaat
     1741 accatcttat tttggcaaca tgactcggtt gcagaagctc catttgaata gcaatagttt
     1801 ccacggaaga atccctcaga gtcttggacg ttgtcgatac ttgctagacc tgtggatgga
     1861 tacaaatagg ttgaatggga ctatacctca ggaaatactg caaattccat ccctcgctta
     1921 catagatttg tcaaacaatt tcttgacagg ccattttcca gaagaagttg gaaagttaga
     1981 acttcttgtt ggactaggtg cttcgtacaa caaattatca ggaaagatgc cacaagctat
     2041 agggggttgt ctttcgatgg aatttctctt tatgcaagga aattcgtttg acggagccat
     2101 tccagatata agccggttgg taagcctaaa gaatgttgac ttctccaaca acaatctctc
     2161 tggccggata cctcgatatc tggccagtct tccttcgctg cgaaatctga atctttctat
     2221 gaacaagttt gagggaaggg tgccaacaac aggagtgttt cgaaatgcta cagcagtttc
     2281 tgtttttggt aacacaaata tttgcggagg cgtccgagaa atgcaactaa agccatgcat
     2341 tgtacaggca tctccaagga agagaaagct ggaggtgctg ttccagggac cagggagcga
     2401 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     2461 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     2521 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     2581 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     2641 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     2701 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     2761 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     2821 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     2881 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     2941 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     3001 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     3061 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     3121 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     3181 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     3241 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3301 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3361 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3421 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3481 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3541 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3601 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4321 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5641 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5701 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5761 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5821 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5881 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5941 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6001 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6061 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
