LOCUS       sequence-296830-        5165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G23300 X009_pECIA2 should be used with prey vector 
            AT2G23300 X009_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5165)
  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 5165)
  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: AT2G23300 X009_pECIA2
FEATURES             Location/Qualifiers
     source          1..5165
                     /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             1418..1441
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2147..2188
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2198..2215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2238..2255)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2288..2422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2312..2331)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2366..2385
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2566..2582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2566..2582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2579..2601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2590..2606
                     /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(2614..2620,2621..2638,2639..2644))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2659..2680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2797..2814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2968..3556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3048..3067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3727..4518,4519..4587))
                     /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     4350..4369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4588..4692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4760..4778
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4816..4838)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4938..4957
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5151..5165,1..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 aactctgatg gagttctctt
      541 actgagtttc aaatactctg ttcttcttga tcctctctct ttattacaat catggaacta
      601 cgaccacgac aatccttgtt catggcgagg tgtgttgtgt aataacgatt caagagttgt
      661 tactttatct ctcccaaact ctaacctcgt tggttcgatt ccttccgatc tgggtttcct
      721 ccaaaacctc caaagtctta atctttccaa taattcactc aatgggtcat taccggttga
      781 gtttttcgcc gccgataagc tccggtttct tgatttatca aataacttga tctccggcga
      841 gatccctgta tcaatcggag gtttacacaa cctccagacg ttaaatctct ccgataacat
      901 cttcaccggg aaactaccag ctaacttagc gtctcttgga agcttaacgg aggtttctct
      961 gaagaacaac tacttctccg gcgagtttcc cggcggcgga tggagatcgg ttcagtatct
     1021 agacatttct tcaaatctaa tcaacggttc actcccacct gatttctccg gcgacaatct
     1081 ccgatacctg aatgtctcgt ataaccaaat ctccggagag attcctccga atgttggtgc
     1141 cggttttcct caaaacgcca ccgttgattt ctccttcaac aatttaaccg gttcaatccc
     1201 agattctccg gtttacctta accagaaatc aatttcgttt tccggaaacc cgggtttatg
     1261 cggaggtccg acccgaaacc cgtgtcccat tccttcatct ccggccaccg tctcgccacc
     1321 aacctctaca cctgcactcg cagctatacc taaatcaatc gggtctaatc gagaaaccga
     1381 accgaacaac aactcaaatc ctcgaaccgg gttaagactg gaggtgctgt tccagggacc
     1441 agggagcgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
     1501 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
     1561 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
     1621 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
     1681 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
     1741 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
     1801 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
     1861 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     1921 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     1981 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     2041 gcaggggaac gtcttctcat gctccgtgat gcatgagggt ctgcacaacc actacacgca
     2101 gaagagcctc tccctgtctc cgggtaaatc tagagggccc ttcgaaggta agcctatccc
     2161 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     2221 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     2281 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2341 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2401 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2461 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     2521 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     2581 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2641 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2701 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2761 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2821 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4741 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4801 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4861 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4921 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4981 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5041 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5101 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     5161 acgtt
//
