LOCUS       sequence-296841-        5195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT4G37250 X020_pECIA2 should be used with prey vector 
            AT4G37250 X020_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5195)
  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 5195)
  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: AT4G37250 X020_pECIA2
FEATURES             Location/Qualifiers
     source          1..5195
                     /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             1448..1471
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2177..2218
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2228..2245
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2268..2285)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2318..2452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2342..2361)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2396..2415
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2596..2612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2596..2612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2609..2631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2620..2636
                     /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(2644..2650,2651..2668,2669..2674))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2689..2710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2827..2844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2998..3586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3078..3097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3757..4548,4549..4617))
                     /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     4380..4399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4618..4722)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4790..4808
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4846..4868)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4968..4987
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5181..5195,1..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 tctgctttaa actctgatgg
      541 gttggttttg atgaagttta aatcctctgt tcttgttgat cctctgtctt tattacagac
      601 atggaattac aaacacgaaa gtccatgttc ttggagagga atctcttgta ataacgattc
      661 taaagtctta actttgtctc ttcccaattc tcagcttctt ggttcgattc cttctgattt
      721 gggttctctc ttaacacttc aaagtctcga tctttccaat aattccttca atgggccatt
      781 accggtttct ttcttcaacg ctagagagct tcgtttcctt gatctctcta gtaacatgat
      841 ctccggcgaa atcccttccg caattggtga tcttcacaat cttctaaccc taaatctctc
      901 tgataatgcc ttggcgggaa agttacctac aaatttggcg tcgttgagaa acttaacggt
      961 ggtttctttg gagaataatt acttttccgg cgagattccc ggcggttgga gagttgttga
     1021 gttcttagac ctgtcgtcga atctgatcaa tggctcgctt cctccggatt ttggcggata
     1081 tagtctccag tacttaaatg tctcgtttaa ccaaatctcc ggcgaaattc caccggagat
     1141 cggagtcaat ttcccaagaa acgtcaccgt tgatctctcc ttcaacaatt taaccggtcc
     1201 aatccctgat tctccggttt tcttaaacca ggaatcaaat ttcttttcag gaaacccggg
     1261 tctatgcggt gaaccgaccc gaaacccgtg tcttatccct tcctcgccgt ctatcgtctc
     1321 cgaagccgac gtaccgacgt ctacaccagc aatcgccgca ataccgaaca caatcggttc
     1381 aaacccggtt accgatccaa attcacaaca aaccgaccca aatccccgaa ccgggttacg
     1441 acccggactg gaggtgctgt tccagggacc agggagcgac aaaactcaca catgcccacc
     1501 gtgcccagca cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa
     1561 ggacaccctc atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca
     1621 cgaagaccct gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa
     1681 gacaaagccg cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt
     1741 cctgcaccag gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct
     1801 cccagccccc atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt
     1861 gtacaccctg cccccatccc gggaggagat gaccaagaac caggtcagcc tgacctgcct
     1921 ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga
     1981 gaacaactac aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag
     2041 caagctcacc gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat
     2101 gcatgagggt ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaatc
     2161 tagagggccc ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     2221 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     2281 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     2341 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2401 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2461 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2521 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     2581 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2641 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2701 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4621 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4681 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4741 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4801 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4861 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4921 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     4981 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5041 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5101 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5161 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
