LOCUS       sequence-276510-        7202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector MIK2 X051_pECIA14 should be used with bait vector MIK2 
            X051_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7202)
  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 7202)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..7202
                     /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             4196..4219
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4235..4252
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4275..4292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4325..4459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4349..4368)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4403..4422
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4603..4619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4603..4619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4616..4638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4627..4643
                     /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(4651..4657,4658..4675,4676..4681))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4696..4717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4834..4851)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5005..5593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5085..5104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5764..6555,6556..6624))
                     /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     6387..6406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6625..6729)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6797..6815
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6853..6875)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6975..6994
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7188..7202,1..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 aagcggcaat cccggcggcg atctggcccc ccagatgctg cgcgagctgc aggagaccaa
     2581 tgccgccctg caggacgtgc gcgagctgct gcgccaccgc gtgaaggaga tcaccttcct
     2641 gaagaacacc gtgatggagt gcgacgcctg cggcatgcag ccctctagag gctccggctc
     2701 catcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     2761 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     2821 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     2881 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     2941 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     3001 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     3061 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     3121 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     3181 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     3241 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     3301 aggtggaggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     3361 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     3421 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     3481 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     3541 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     3601 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     3661 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     3721 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     3781 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     3841 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     3901 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     3961 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     4021 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     4081 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     4141 cggcaccacc gacgccgcgc acccgggttc tagaggaagc acctccggcg ataacgatta
     4201 caaggatgat gatgataagg ccagcggcac cggtcatcat caccatcacc attgagttta
     4261 aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat cagcctcgat
     4321 cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4381 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4441 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4501 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     4561 tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     4621 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4681 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4741 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4801 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4861 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4921 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4981 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5041 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5101 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5161 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5221 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5281 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5341 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5401 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5461 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5521 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5581 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5641 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5701 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5761 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5821 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5881 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5941 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6001 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6061 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6121 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6181 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6241 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6301 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6361 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6421 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6481 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6541 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6601 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6661 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6721 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6781 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6841 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6901 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6961 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7021 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     7081 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     7141 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7201 tt
//
