LOCUS       sequence-270265-        5915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector PRK8 I11_pECIA14 should be used with bait vector PRK8 
            I11_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5915)
  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 5915)
  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: PRK8 I11_pECIA14
FEATURES             Location/Qualifiers
     source          1..5915
                     /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             1211..1234
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2909..2932
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2948..2965
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2988..3005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3038..3172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3062..3081)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3116..3135
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3316..3332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3316..3332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3329..3351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3340..3356
                     /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(3364..3370,3371..3388,3389..3394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3409..3430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3547..3564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3718..4306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3798..3817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4477..5268,5269..5337))
                     /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     5100..5119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5338..5442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5510..5528
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5566..5588)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5688..5707
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5901..5915,1..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 tcgataaccg aatcagaatc
      541 tttattgaaa ttcaagaagt ccttaaacaa caccaagtct cttgattctt ggacaccaga
      601 gtcagaacct tgtggagcaa gccaacgatg gatcggatta ctctgcaaca aaaactctgt
      661 ctttggcctc cagatcgaac aaatgggtct atcaggaaaa gttgatgtcg ctccgttaaa
      721 agacctacca tctttacgca ccataagcat aatgaacaac tctttctctg gtgacatacc
      781 cgagtttaac agattaaccg ctctgaagtc gctttacata tcaggaaatc gattctctgg
      841 taacattcct tctgattatt tcgaaacaat ggtgtcgcta aagaaagctt ggctgtcaaa
      901 caaccatttc tctggtctca ttcctatctc attggcaact acattgccaa atctcattga
      961 gttacgcctc gaaaataatc aattcatcgg gagtatacct aatttcacac aaacgacttt
     1021 agctattgtt gatctatcaa acaaccaact aaccggagaa atacctcccg gtttgttgaa
     1081 attcgacgcg aagacttttg cggggaattc tggcctttgc ggtgccaagc tatccactcc
     1141 atgcccacaa ccaaaaaatt cgactgcatc cataacgata gaaggaacaa tgaaagatgc
     1201 aaataagagc ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
     1261 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
     1321 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
     1381 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
     1441 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     1501 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     1561 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     1621 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     1681 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     1741 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     1801 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     1861 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     1921 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     1981 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
     2041 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     2101 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
     2161 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     2221 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     2281 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     2341 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     2401 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     2461 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     2521 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     2581 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     2641 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     2701 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     2761 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     2821 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     2881 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
     2941 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     3001 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     3061 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3121 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3181 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3241 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     3301 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3361 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3421 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3481 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3541 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3601 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3661 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3721 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4141 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4321 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4381 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4441 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4501 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4561 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4621 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4681 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4741 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4801 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4861 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4921 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4981 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5041 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5101 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5161 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5221 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5281 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5401 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5461 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5521 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5581 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5641 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5701 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5761 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5821 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5881 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
