LOCUS       sequence-297007-        5015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G41180 P3_pECIA2 should be used with prey vector 
            AT5G41180 P3_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5015)
  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 5015)
  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: AT5G41180 P3_pECIA2
FEATURES             Location/Qualifiers
     source          1..5015
                     /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             1268..1291
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1997..2038
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2048..2065
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2088..2105)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2138..2272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2162..2181)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2216..2235
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2416..2432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2416..2432)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2429..2451)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2440..2456
                     /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(2464..2470,2471..2488,2489..2494))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2509..2530
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2647..2664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2818..3406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2898..2917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3577..4368,4369..4437))
                     /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     4200..4219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4438..4542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4610..4628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4666..4688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4788..4807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5001..5015,1..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 tctaaagaag ttgaagcagt
      541 tagaagattc aaggaagcca tttataagga cccattgcta gttatgtcta attggaatgt
      601 ccccaatttg agtccttgtg attggaatgg cattaaatgt tctccatcta aggatcacat
      661 tatcaagata aatatatcgg ggacatcgat gagagggttt cttgtgccag aacttggtca
      721 aataacctac ttgcaagaac tgatcctgcg tgggaacatt ctaatgggga caataccaaa
      781 ggagatagga aagttaaaga aactcaagat cttagacctg ggaaacaatc atttgacagg
      841 accgattcca gcagagatcg ggaaattgtc aaggattaag acaataaacc ttcagtccaa
      901 tggtttaata ggaaagttac ctccagagat tggaaacttg aagcacctta aagaacttct
      961 tattggcagg aataggcttc gaggaagtat tcctattgcc gcgaaaacat caaaaaagta
     1021 tgcttcaaat ccaagtgcaa acatcagtgg tttgtgcaag tcttctctat ttaaagtggc
     1081 agatttctct tacaactttt tcgagggaag agttccgagt tgcttggatt acctcccaat
     1141 aacgagcttt caaggaaact gcatgaaaac catggatgtt aagcagagac ctctttcaga
     1201 atgtgctcgc ttagctgtaa ccgtggccaa gaagaagcat cgagcatcga gacaaacatg
     1261 gcttcggctg gaggtgctgt tccagggacc agggagcgac aaaactcaca catgcccacc
     1321 gtgcccagca cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa
     1381 ggacaccctc atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca
     1441 cgaagaccct gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa
     1501 gacaaagccg cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt
     1561 cctgcaccag gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct
     1621 cccagccccc atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt
     1681 gtacaccctg cccccatccc gggaggagat gaccaagaac caggtcagcc tgacctgcct
     1741 ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga
     1801 gaacaactac aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag
     1861 caagctcacc gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat
     1921 gcatgagggt ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaatc
     1981 tagagggccc ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     2041 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     2101 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     2161 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2221 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2281 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2341 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     2401 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2461 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2521 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3241 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4561 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4621 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4681 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4741 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4801 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     4861 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4921 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     4981 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
