LOCUS       sequence-296775-        4958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector PRK8 I11_pECIA2 should be used with prey vector PRK8 
            I11_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4958)
  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 4958)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..4958
                     /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             1940..1981
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1991..2008
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2031..2048)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2081..2215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2105..2124)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2159..2178
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2359..2375)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2359..2375)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2372..2394)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2383..2399
                     /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(2407..2413,2414..2431,2432..2437))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2452..2473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2590..2607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2761..3349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2841..2860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3520..4311,4312..4380))
                     /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     4143..4162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4381..4485)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4553..4571
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4609..4631)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4731..4750
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4944..4958,1..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 accagggagc gacaaaactc acacatgccc
     1261 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     1321 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     1381 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     1441 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     1501 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     1561 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     1621 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     1681 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     1741 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     1801 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     1861 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     1921 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     1981 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2041 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     2101 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2161 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2221 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2281 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     2341 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2401 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2461 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2521 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2581 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2641 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3781 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3961 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4261 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4381 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4441 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4501 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4561 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4621 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4681 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4741 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4801 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4861 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4921 aagttgggta acgccagggt tttcccagtc acgacgtt
//
