LOCUS       sequence-276506-        7001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector IKU2 X076_pECIA14 should be used with bait vector IKU2 
            X076_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7001)
  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 7001)
  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: IKU2 X076_pECIA14
FEATURES             Location/Qualifiers
     source          1..7001
                     /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             2297..2320
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3995..4018
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4034..4051
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4074..4091)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4124..4258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4148..4167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4202..4221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4402..4418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4402..4418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4415..4437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4426..4442
                     /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(4450..4456,4457..4474,4475..4480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4495..4516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4633..4650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4804..5392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4884..4903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5563..6354,6355..6423))
                     /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     6186..6205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6424..6528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6596..6614
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6652..6674)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6774..6793
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6987..7001,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
       61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactcaggcc
      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 tccaaccaca gtgaagaagt
      541 cgagaatctt ctcaaactga aatccacctt tggagaaaca aaatccgacg acgtctttaa
      601 gacctggact caccggaatt cggcatgtga attcgccgga atcgtatgta attccgacgg
      661 aaacgtcgtc gagatcaacc ttggaagtcg gagtttgatc aaccgagacg acgacggtag
      721 gttcaccgat cttcctttcg attcaatctg cgatttgaag ttgctggaga agcttgttct
      781 aggaaacaac tctttacgtg gtcagattgg tacgaatctc gggaaatgta atcgcttgag
      841 atacttagac ctcgggatca acaatttctc cggtgagttt ccggcgattg attcgttaca
      901 attactcgag tttttgagct taaacgcttc tggaatctct ggaatattcc catggagttc
      961 tctaaaagac ctgaaacgat tgagtttcct aagcgttgga gataaccgat tcggttcgca
     1021 tccatttcct agagagattc taaatctcac tgccttgcaa tgggtttatc tgagtaacag
     1081 cagcatcact ggtaagattc cagagggaat caaaaacctt gttcgtctac aaaacttgga
     1141 gctctctgat aatcagatct ctggtgaaat tccaaaggag attgttcagc tcaaaaacct
     1201 aaggcagctt gagatttata gtaatgactt gaccggtaag ttaccattgg gttttaggaa
     1261 tttgacgaat ctgaggaatt tcgacgcctc taacaactcc ttggaaggtg atttatcaga
     1321 gctgcggttc ttaaagaatc ttgtgtctct tggaatgttc gaaaaccgat taaccggtga
     1381 gattccaaaa gagtttggag atttcaagag tttagctgct ttgtctcttt acagaaacca
     1441 gctcacaggg aagcttccga ggagactcgg ttcttggaca gcttttaagt acatcgacgt
     1501 gtctgagaac ttcttggaag gtcagattcc tccttatatg tgcaagaaag gcgtaatgac
     1561 gcatcttctc atgttgcaga acagattcac aggacaattt ccagagagtt atgctaaatg
     1621 taagactctg attcgtcttc gtgtaagtaa caactctctc tctggaatga ttccttcagg
     1681 gatttggggt cttcctaatc tacagtttct tgaccttgct tcaaactatt ttgaaggaaa
     1741 cctcactggt gatattggta atgccaagtc tctaggttct ttagatttga gcaataatag
     1801 attttcaggt tctctaccgt ttcagatctc aggagctaac tcgttagtgt cggttaatct
     1861 tcgaatgaat aagttttccg ggatagttcc tgaatctttt ggtaagctta aggagctttc
     1921 aagtctcatt cttgaccaga ataatctatc tggtgctata ccaaagtcac taggcttatg
     1981 cacatctttg gttgatctaa actttgctgg aaactcactc tctgaggaaa tcccggaaag
     2041 tttaggatct ttaaagttat tgaattccct gaatctgtca ggaaacaaat tatcaggaat
     2101 gattcctgtt ggtttgtcgg ctctgaagct aagcttactt gacttatcca acaaccaatt
     2161 gaccggttct gttcctgaat ctctcgtaag tggaagcttt gaaggaaatt caggactatg
     2221 cagctcaaag atcagatatc ttcgtccatg tccgcttgga aaacctcaca gccaagggaa
     2281 gaggaaacat ttgtcactgg aggtgctgtt ccagggacca agcggcaatc ccggcggcga
     2341 tctggccccc cagatgctgc gcgagctgca ggagaccaat gccgccctgc aggacgtgcg
     2401 cgagctgctg cgccaccgcg tgaaggagat caccttcctg aagaacaccg tgatggagtg
     2461 cgacgcctgc ggcatgcagc cctctagagg ctccggctcc atcatcccag ttgaggagga
     2521 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     2581 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     2641 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     2701 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     2761 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     2821 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     2881 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     2941 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     3001 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     3061 tacgcagctc atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat
     3121 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     3181 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     3241 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     3301 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     3361 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     3421 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     3481 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     3541 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     3601 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     3661 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     3721 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     3781 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     3841 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     3901 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     3961 cccgggttct agaggaagca cctccggcga taacgattac aaggatgatg atgataaggc
     4021 cagcggcacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     4081 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     4141 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4201 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4261 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4321 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga cctgcaggca
     4381 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6541 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6601 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6661 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6721 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6781 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6841 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6901 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6961 attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
