LOCUS       sequence-297039-        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G56140 X032_pECIA2 should be used with prey vector 
            AT1G56140 X032_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  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 6071)
  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: AT1G56140 X032_pECIA2
FEATURES             Location/Qualifiers
     source          1..6071
                     /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             2324..2347
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3053..3094
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3104..3121
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3144..3161)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3194..3328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3272..3291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3472..3488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3485..3507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3496..3512
                     /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(3520..3526,3527..3544,3545..3550))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3565..3586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3703..3720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3874..4462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3954..3973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4633..5424,5425..5493))
                     /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     5256..5275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5494..5598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5666..5684
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5722..5744)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5844..5863
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6057..6071,1..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 acaggagcca ccacacatcc
      541 cgacgaagcg ctagctttga actcaatttt cgcggcttgg aggattcggg cgccgaggga
      601 atggaacatc agcggcgaac tttgctccgg cgccgctatc gacgccagtg ttctcgactc
      661 aaaccctgcc tacaatcctc tcatcaaatg cgactgcagt ttcgaaaact ccacaatctg
      721 ccgcattacc aacatcaagg tttatgcgat ggaagttgta ggatctatac ctcaacaact
      781 ctggaccttg gaatacctga caaatctgaa cttgggtcaa aatgttctca ctggctcact
      841 tcctcctgca cttggaaatt tgactcgaat gcgatggatg acttttggga tcaatgcgtt
      901 gtctggccct attcctaaag aaatcggttt gcttacagat ttaagattac ttagtattag
      961 ttcaaataac ttttcgggtt ctataccaga tgagattggg agatgtacaa aactacaaca
     1021 gatatacata gatagttcgg gactcagcgg gggattacct gtatcatttg ctaatcttgt
     1081 ggagctggaa caagcttgga ttgcggatat ggaacttaca ggtcagatac cagactttat
     1141 aggagattgg accaaactta ctaccttgag aattcttgga actggtttga gtggtccgat
     1201 accggcgtcg ttttcaaact taacttcttt gacagagctg aggcttggtg atatatccaa
     1261 tggaaattct tctcttgaat tcatcaaaga catgaaatct ctaagtatat tagtattgag
     1321 gaacaataat ctcactggga caataccgtc taatattgga gaatattcaa gtttgcgaca
     1381 acttgatttg agcttcaaca aactacatgg aacaattccg gcttcacttt tcaacttaag
     1441 acagcttact cacttgtttc tggggaacaa cacgttgaat ggctcattgc ccactcaaaa
     1501 ggggcagtct ctgagcaatg tagatgtgtc atacaatgat ttgtctggaa gtcttccgtc
     1561 atgggtcagt ttaccaaact tgaatctcaa cttagttgct aacaacttca cattggaagg
     1621 tcttgacaac agggttttat caggactgaa ctgcctgcag aagaacttcc catgcaatcg
     1681 agggaaagga atatattctg acttctcaat caactgcgga ggcccagaga taagatctgt
     1741 aactgaagca gtatttgaga gggaggacga ggatcttgga ccagcttcat ttgtcgtgag
     1801 tgctggtcag agatgggcag ccagtagtgt aggacttttt gccggaagta gcaacaatat
     1861 atacatatct acttcacaat cacaatttgt caacactttg gactcagagc tatttcagtc
     1921 agcaagactt tctgcatctt ccctaaggta ttatgggttg ggtctagaaa atggaggcta
     1981 taccgtcaca cttcagtttg ctgaaataca aattctaggt tctacttcca acacttggag
     2041 aggtttagga agacgacgtt ttgacattta tgtccaggga agacttgttg aaaaggactt
     2101 cgatgtacgc agaacagctg gtgactccac tgttcgagca gttcagagag aatataaagc
     2161 aaatgtatca caaaatcatc tcgaaattca tcttttctgg gctggaaaag gaacatgctg
     2221 tattcctatc caaggggctt atgggccatt aatatcggcc gtcggtgcaa caccagattt
     2281 cacaccaact gtgggtaata ggccaccatc aaagggaaag agcctggagg tgctgttcca
     2341 gggaccaggg agcgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg
     2401 gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac
     2461 ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa
     2521 ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta
     2581 caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg
     2641 caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
     2701 ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga
     2761 ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga
     2821 catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc
     2881 cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag
     2941 gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gagggtctgc acaaccacta
     3001 cacgcagaag agcctctccc tgtctccggg taaatctaga gggcccttcg aaggtaagcc
     3061 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     3121 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     3181 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     3241 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3301 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3361 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3421 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     3481 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3541 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3601 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3661 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3721 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3781 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3841 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3901 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3961 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4021 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4081 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4141 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4201 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4261 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4321 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4381 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4501 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5641 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5701 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5761 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5821 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5881 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     5941 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6001 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6061 gtcacgacgt t
//
