LOCUS       sequence-298824-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector MIK2 X051_pECIA2 should be used with prey vector MIK2 
            X051_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  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 6245)
  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: MIK2 X051_pECIA2
FEATURES             Location/Qualifiers
     source          1..6245
                     /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             2498..2521
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3227..3268
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3278..3295
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3318..3335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3368..3502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3392..3411)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3446..3465
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3646..3662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3659..3681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3670..3686
                     /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(3694..3700,3701..3718,3719..3724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3877..3894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4048..4636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4128..4147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4807..5598,5599..5667))
                     /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     5430..5449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5668..5772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5840..5858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5896..5918)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6018..6037
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6231..6245,1..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 tctgcaacgg tggaagaagc
      541 taatgctctt ttgaaatgga agtcaacttt cacaaaccag acaagttcat caaagctatc
      601 ttcttgggtc aatccaaaca caagcagctt ttgtaccagt tggtatggtg ttgcctgctc
      661 actcgggagc attatacggt tgaatcttac taataccggt atcgaaggta ctttcgaaga
      721 ttttccattc tcgtctctcc ccaatctcac ttttgttgat ctcagtatga accgtttctc
      781 tggaaccatc tctcctttat ggggacggtt ctcgaaactc gaatacttcg atttgtcgat
      841 taaccagtta gttggtgaaa tcccacctga acttggtgat ttgagtaatt tagatactct
      901 ccatctagtt gaaaacaagt taaacggatc gattccttct gagatcggtc ggttaactaa
      961 ggttacggag atagctatat acgacaatct tttaaccgga ccaatacctt cttcttttgg
     1021 aaaccttacc aaactagtta acctctacct ctttatcaat tctctttctg gttcaattcc
     1081 ttctgaaata ggaaacttac ccaaccttag agagctatgt ctagatagga acaaccttac
     1141 cggtaaaatc ccttctagtt tcgggaattt gaagaatgta actcttctca atatgtttga
     1201 aaatcagctc tctggtgaaa tccctcctga gattggtaat atgaccgctt tagatacact
     1261 tagtctccac acaaataagc ttaccggtcc aataccttct accttaggaa acatcaaaac
     1321 cctagccgtt cttcatcttt acctgaatca actcaatggt tcgattccac cggaactagg
     1381 cgaaatggaa tcaatgattg atttggagat aagtgagaac aaactcaccg gtcctgttcc
     1441 cgattctttt ggcaagttaa ccgcgttgga atggttgttc ctacgtgata atcagctctc
     1501 tggtcccatc ccaccgggaa ttgctaactc tacagagctc accgtcctgc agctcgacac
     1561 caacaacttc accggtttct tgcctgatac catctgtagg ggtggaaagc ttgagaatct
     1621 tacgctagat gataatcact tcgaaggtcc tgttcctaaa agcttgagag attgcaagag
     1681 cttgatcaga gtaagattca aagggaatag tttctccgga gatatctctg aggctttcgg
     1741 agtttacccg actctcaact tcatcgatct cagcaacaac aactttcatg gacaactttc
     1801 agccaactgg gaacaaagtc aaaagctggt tgcattcatc ctctcaaaca acagcatcac
     1861 tggagccatc ccacctgaga tttggaacat gacacagcta agtcagctcg atctatcttc
     1921 caaccgcatc accggcgagc tacctgaatc aatcagcaat atcaaccgta tatcaaaact
     1981 ccaactcaac gggaatcggc tatcaggcaa aatcccttca ggaatcagac tcttaaccaa
     2041 tcttgagtat cttgatctat cctcaaaccg attcagttcc gaaatacccc caacgctcaa
     2101 caatttgcca aggctctact acatgaacct gagcagaaac gacttggatc aaaccatccc
     2161 ggaggggtta acaaagctat ctcagttaca aatgcttgat ctcagctaca atcagctcga
     2221 cggagagatc tcatcacaat tcagatcttt acaaaacctc gaaagactcg acctctcaca
     2281 caacaatctc tcaggtcaaa tcccaccaag tttcaaagat atgctagcac taacacacgt
     2341 cgatgtatca cacaacaacc tccaaggtcc aattccagat aacgcagcgt ttcgaaacgc
     2401 acctccagat gcttttgaag gtaacaaaga cttatgtggt agtgttaaca ccacacaagg
     2461 cttgaagcca tgttcaatca catcctcaaa gaaatcactg gaggtgctgt tccagggacc
     2521 agggagcgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
     2581 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
     2641 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
     2701 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
     2761 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
     2821 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
     2881 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
     2941 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     3001 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     3061 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     3121 gcaggggaac gtcttctcat gctccgtgat gcatgagggt ctgcacaacc actacacgca
     3181 gaagagcctc tccctgtctc cgggtaaatc tagagggccc ttcgaaggta agcctatccc
     3241 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     3301 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     3361 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3421 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3481 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3541 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     3601 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5821 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5881 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5941 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6001 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6061 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     6121 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6181 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6241 acgtt
//
