LOCUS       sequence-296782-        4985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector PRK4 J6_pECIA2 should be used with prey vector PRK4 
            J6_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4985)
  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 4985)
  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: PRK4 J6_pECIA2
FEATURES             Location/Qualifiers
     source          1..4985
                     /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             1238..1261
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1967..2008
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2018..2035
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2058..2075)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2108..2242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2132..2151)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2186..2205
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2386..2402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2386..2402)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2399..2421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2410..2426
                     /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(2434..2440,2441..2458,2459..2464))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2479..2500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2617..2634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2788..3376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2868..2887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3547..4338,4339..4407))
                     /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     4170..4189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4408..4512)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4580..4598
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4636..4658)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4758..4777
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4971..4985,1..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 tctcagccgc aggcggatgt
      541 tttgccactg ccagcctcag acgcagattg cctcttgaga tttaaagata ctttggttaa
      601 tgcatcgttc attagcagtt gggatccttc catctccccg tgtaagcgaa actcagagaa
      661 ttggttcggt gttctctgtg ttaccggtaa tgtttggggc ctacaactcg aaggaatggg
      721 cttaaccggg aagcttgacc ttgaaccatt agctgcaatc aagaatctac gaaccttgag
      781 cttcatgaac aacaaattta acggttcaat gccatctgtc aagaattttg gtgcgttgaa
      841 atcattgtac ttgtctaaca accggtttac aggggagata cccgcggatg cgtttgatgg
      901 tatgcatcat ttgaagaagc ttctgttggc taacaacgcg tttcgaggga gtatcccttc
      961 ttctttagct tatttgccaa tgcttttaga gttgaggcta aatgggaatc agtttcatgg
     1021 ggaaatacct tattttaaac aaaaggacct taagttggct agcttcgaaa acaatgacct
     1081 cgagggacct ataccggaaa gccttagcaa catggatcct gtctcctttt cagggaacaa
     1141 gaacttgtgt ggtcctccac taagcccatg ttcgagtgat tcaggatcgt ctccggatct
     1201 cccttctagt cccacggaaa agaacaagaa ccaatctctg gaggtgctgt tccagggacc
     1261 agggagcgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
     1321 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
     1381 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
     1441 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
     1501 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
     1561 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
     1621 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
     1681 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     1741 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     1801 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     1861 gcaggggaac gtcttctcat gctccgtgat gcatgagggt ctgcacaacc actacacgca
     1921 gaagagcctc tccctgtctc cgggtaaatc tagagggccc ttcgaaggta agcctatccc
     1981 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     2041 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     2101 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2161 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2221 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2281 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     2341 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     2401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4561 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4621 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4681 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4741 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     4801 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     4861 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     4921 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     4981 acgtt
//
