LOCUS       sequence-296497-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G51810 M2_pECIA2 should be used with prey vector 
            AT1G51810 M2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  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 5411)
  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: AT1G51810 M2_pECIA2
FEATURES             Location/Qualifiers
     source          1..5411
                     /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             1664..1687
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2393..2434
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2444..2461
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2484..2501)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2534..2668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2558..2577)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2612..2631
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2812..2828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2812..2828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2825..2847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2836..2852
                     /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(2860..2866,2867..2884,2885..2890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2905..2926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3043..3060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3214..3802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3294..3313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3973..4764,4765..4833))
                     /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     4596..4615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4834..4938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5006..5024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5062..5084)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5184..5203
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5397..5411,1..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 atgtggatca cggtgaatac
      541 tgataacact atcaaggaga tcctccacgt atcgaaatca aacactttgc aagtgtgtct
      601 tgttaagaca ggtacaagta taccttatat aaatacattg gaactacgac cattggccga
      661 cgatatatac accaacgaaa gtggctccct caactatctt tttcgggttt attatagcaa
      721 tttaaagggc tatatagagt accccgatga tgtccacgat cgcatatgga aacaaatcct
      781 accttaccag gattggcaga ttttaactac gaatctccaa ataaacgttt ctaatgatta
      841 tgatctaccc caacgtgtaa tgaaaacagc tgtaacaccc attaaagcta gcacaacgac
      901 gatggaattt ccctggaact tagagcctcc aacttcacag ttttacttat tccttcactt
      961 tgcagagctt caaagtctac aagccaacga gacgagggaa ttcaatgtgg tgttgaacgg
     1021 aaatgttaca tttaaatctt atagtcctaa gtttttagaa atgcaaacag tatatagcac
     1081 agcaccaaag caatgcgatg gagggaaatg cttgttgcag ttagtgaaaa cgtcaaggtc
     1141 cactttgccg cctctaatta atgctatgga ggcttacact gtgcttgatt tcccacagat
     1201 agaaacaaat gtagatgaag tgattgctat caagaatata caatctactt atggattgag
     1261 taaaacaacc tggcaaggag atccatgtgt acccaaaaag ttcttgtggg atggtttaaa
     1321 ctgcaacaac tcggatgatt ctacgccacc aattatcact tccttaaact tgtcttcaag
     1381 tggattgact ggaattatcg tgctgaccat tcagaatctc gccaatttac aagaactgga
     1441 cttgtcaaat aacaacttgt ctggaggtgt tcctgaattt ctagcggata tgaagtcgct
     1501 cttggtcata aacttaagtg ggaacaatct cagtggtgta gttcctcaaa agcttataga
     1561 gaagaaaatg ttgaaattga acattgaagg caatccgaag cttaattgca cagtggagtc
     1621 atgtgtaaac aaagatgaag agggtggacg acagataaag agcctggagg tgctgttcca
     1681 gggaccaggg agcgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg
     1741 gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac
     1801 ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa
     1861 ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta
     1921 caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg
     1981 caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
     2041 ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga
     2101 ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga
     2161 catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc
     2221 cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag
     2281 gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gagggtctgc acaaccacta
     2341 cacgcagaag agcctctccc tgtctccggg taaatctaga gggcccttcg aaggtaagcc
     2401 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     2461 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     2521 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     2581 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2641 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2701 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     2761 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     2821 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2881 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2941 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3001 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4981 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5041 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5101 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5161 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5221 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     5281 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5341 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5401 gtcacgacgt t
//
