LOCUS       sequence-238701-        7541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RPK2 X042_pECIA14 should be used with bait vector RPK2 
            X042_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7541)
  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 7541)
  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: RPK2 X042_pECIA14
FEATURES             Location/Qualifiers
     source          1..7541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             3282..3305
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4980..5003
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5019..5036
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5059..5076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5109..5243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5133..5152)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5187..5206
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5387..5403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5387..5403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5400..5422)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5411..5427
                     /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(5435..5441,5442..5459,5460..5465))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5480..5501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5618..5635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5789..6377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5869..5888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6548..7339,7340..7408))
                     /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     7171..7190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7409..7513)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggaaaat cactgtttta gctgattcgg acaaatccgt gttgcttcgg tttaagaaaa
     1021 cggtatcgga ccctggttcg attctagcta gctgggtcga agagagtgaa gattactgct
     1081 cctggtttgg tgtatcttgc gattcgagct ctcgagtcat ggctcttaac attagtggct
     1141 ctggaagttc tgaaattagt cgaaatcgtt tcacttgtgg tgatattggt aagtttccgt
     1201 tgtatgggtt tggtgttcgg agggattgta ccggaaatca tggagctttg gcggggaatt
     1261 tgccttctgt aatcatgagt ctcacaggac ttagggtttt gtctctgcct tttaattcgt
     1321 ttagtggaga gattcctgta ggaatctggg gaatggagaa gcttgaagtt cttgatctgg
     1381 aagggaattt gatgacgggg tcattgccgg atcagtttac tgggttacgg aatttgcggg
     1441 ttatgaattt ggggtttaac agagtttcag gtgaaatccc taattcgctg cagaatctga
     1501 caaagttgga gattttgaat ttgggtggta ataagttgaa tggaactgtt cctggttttg
     1561 ttgggagatt cagagtgttg catttgccgt taaactggct tcaaggctct ttgcctaaag
     1621 atattgggga tagctgtgga aagcttgagc atttagattt gtctggtaat ttcttgactg
     1681 ggaggattcc agagagtttg gggaaatgtg ctggtttaag gtcgttgttg ttgtatatga
     1741 atacgttgga ggagactatc cctttagaat ttgggagtct tcagaagctt gaggttttgg
     1801 atgtttctag gaacactctt agtggtccat tacctgttga acttggaaat tgctcttcgt
     1861 tgtctgttct cgttctatca aatctgtaca atgtgtatga agacattaat agtgtaagag
     1921 gggaagctga tcttcctcca ggcgctgatt taacctcgat gaccgaggat ttcaatttct
     1981 atcaaggagg aatcccggag gaaatcacta ggcttccgaa actaaagata ctttgggtgc
     2041 ccagagctac gttagaaggt agatttccag gagattgggg ttcatgtcaa aacttggaga
     2101 tggttaattt gggtcagaac ttctttaaag gcgaaattcc tgtcggtctt agtaaatgca
     2161 agaaccttcg tcttcttgat ttgagctcga ataggcttac tggggagctt ctcaaagaga
     2221 tttcggtccc atgcatgagt gtttttgatg ttggtggaaa cagcttatcg ggtgtaatcc
     2281 ctgattttct caacaatacc acaagccatt gtcctcctgt tgtttacttt gacaggtttt
     2341 ctattgaatc ttacagcgac ccgtcgtctg tttatctctc cttctttacc gagaaggctc
     2401 aagttggaac ttccctgata gatcttggaa gtgatggagg tccggccgtg tttcataatt
     2461 ttgcagacaa caacttcact ggtactctca agtctatacc acttgcacag gagcggttgg
     2521 gaaagcgagt ctcttacata ttttctgctg gagggaatcg gttgtacggg caatttccgg
     2581 gaaatctgtt tgataactgc gacgaactaa aagctgttta tgtcaatgta agcttcaaca
     2641 agctctcagg tcggattcct cagggactaa acaacatgtg cacttctctt aagattcttg
     2701 atgcctcggt gaatcagatc ttcgggccaa tcccaacaag tctcggggat ctagcttcac
     2761 ttgttgctct taatctgagc tggaatcagt tgcaaggtca gataccgggt agcctcggga
     2821 agaagatggc agctttgaca tatctttcga ttgcgaataa caacctcacg gggcaaattc
     2881 ctcagagctt tggtcagtta cattccttgg atgttcttga tctctcttca aatcatctat
     2941 ctggcggtat tcctcatgat ttcgtaaact tgaagaatct cactgtcctg cttctcaaca
     3001 ataacaacct ctctggccca atcccgtcgg gttttgcgac atttgctgtt ttcaatgttt
     3061 cctccaacaa tctgtctggt ccagttcctt caaccaatgg actgacgaaa tgcagcactg
     3121 ttagtggaaa cccctatctt cgaccttgcc atgtgttttc tctgacaaca ccgtcttcgg
     3181 attcccgaga ctccacgggt gattctatca cgcaggatta cgcatcttcg cctgttgaaa
     3241 acgctccatc tcagtcgcca ggaaagggtg gattcaactc gctggaggtg ctgttccagg
     3301 gaccaagcgg caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga
     3361 ccaatgccgc cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct
     3421 tcctgaagaa caccgtgatg gagtgcgacg cctgcggcat gcagccctct agaggctccg
     3481 gctccatcat cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg
     3541 ccctgggtgc cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct
     3601 tcctgggcga tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga
     3661 agaaggacaa actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc
     3721 tgtccaagac atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct
     3781 acctgtgcgg ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta
     3841 accagtgcaa cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag
     3901 cagggaagtc agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca
     3961 cctacgccca cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc
     4021 gccaggaggg gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga
     4081 tcctaggtgg aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag
     4141 atgactacag ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg
     4201 cgaagcgcca gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg
     4261 acccgtctgt gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc
     4321 accgagactc cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc
     4381 tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc
     4441 atcatgaaag cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg
     4501 agagggcggg ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact
     4561 cccacgtctt ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc
     4621 ctggcaaggc ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct
     4681 atgtgctcaa ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt
     4741 atcggcagca gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg
     4801 tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag
     4861 cgcacgtcat ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc
     4921 ccgccggcac caccgacgcc gcgcacccgg gttctagagg aagcacctcc ggcgataacg
     4981 attacaagga tgatgatgat aaggccagcg gcaccggtca tcatcaccat caccattgag
     5041 tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct
     5101 cgatcgagga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5161 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5221 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     5281 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     5341 attatgatca gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc
     5401 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5461 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5521 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5581 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6841 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7141 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7501 taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
