LOCUS       sequence-238711-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RPK2 X042_pECIA2 should be used with prey vector RPK2 
            X042_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6584
                     /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             4011..4052
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4062..4079
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4102..4119)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4152..4286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4176..4195)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4230..4249
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4430..4446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4430..4446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4443..4465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4454..4470
                     /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(4478..4484,4485..4502,4503..4508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4523..4544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4661..4678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4832..5420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4912..4931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5591..6382,6383..6451))
                     /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     6214..6233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6452..6556)
                     /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 gacgaactca 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 gaccagggag cgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg
     3361 gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc
     3421 ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact
     3481 ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca
     3541 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca
     3601 aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct
     3661 ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg
     3721 agatgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca
     3781 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg
     3841 tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt
     3901 ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga gggtctgcac aaccactaca
     3961 cgcagaagag cctctccctg tctccgggta aatctagagg gcccttcgaa ggtaagccta
     4021 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt
     4081 gagtttaaac ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag
     4141 cctcgatcga ggatccagac atgataagat acattgatga gtttggacaa accacaacta
     4201 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     4261 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     4321 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     4381 ctgattatga tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt
     4441 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4501 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4621 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4681 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4741 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4801 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4861 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4921 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4981 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5041 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5101 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5161 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5221 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5281 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5341 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5401 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5461 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5521 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5581 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5641 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5701 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5761 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5821 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5881 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5941 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6001 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6061 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6121 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6181 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6241 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6301 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6361 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6421 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6481 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6541 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgac
//
