LOCUS       sequence-298773-        6161 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector PSKR1 nC5_pECIA2 should be used with prey vector PSKR1 
            nC5_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6161)
  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 6161)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: PSKR1 nC5_pECIA2
FEATURES             Location/Qualifiers
     source          1..6161
                     /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             2414..2437
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3143..3184
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3194..3211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3234..3251)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3284..3418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3308..3327)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3362..3381
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3562..3578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3562..3578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3575..3597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3586..3602
                     /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(3610..3616,3617..3634,3635..3640))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3655..3676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3793..3810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3964..4552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4044..4063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4723..5514,5515..5583))
                     /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     5346..5365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5584..5688)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5756..5774
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5812..5834)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5934..5953
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6147..6161,1..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 tcggaatctc agaccacctc
      541 caggtgccat ccacatgacc tcgaagcctt acgtgacttc atagcacatc tcgaaccaaa
      601 accagatggt tggatcaatt cttcttcttc tacagactgc tgcaactgga ccggaatcac
      661 ctgcaattca aacaacaccg gaagagttat tagattggag cttgggaaca aaaagctgtc
      721 ggggaagttg tctgaatctc tcgggaagct agatgagatt agggttctta atctctctcg
      781 aaacttcatc aaagattcga tccctctttc gattttcaac ttgaagaatc tacaaactct
      841 tgatttgagc tctaatgatc tctccggcgg aatcccaaca agtataaatc tcccagctct
      901 gcaaagtttt gatctttctt caaataaatt caatgggtcg cttccgtctc atatctgcca
      961 taactctact caaattaggg ttgtgaaact tgcggtgaac tacttcgccg gaaacttcac
     1021 ttccgggttt gggaaatgtg tcttgcttga gcatctctgt cttggtatga acgatcttac
     1081 tggtaacatc cctgaggatt tgtttcatct caaaagattg aatcttttag ggattcaaga
     1141 gaatcgtctc tctggttcgt tgagtcgtga gattaggaat ctctcaagtc ttgttcgtct
     1201 tgatgtttct tggaatttgt tttccggtga aatccctgat gtgttcgacg aattgcctca
     1261 gttaaagttt ttcttaggtc agaccaatgg attcattgga ggaataccta aatcgttggc
     1321 gaattcaccg agtttgaatc tgcttaactt gaggaacaat tctttatcgg gtcgtttgat
     1381 gttgaattgt acggcgatga ttgctttgaa ctctcttgat ttaggtacca atagattcaa
     1441 tgggaggtta cctgagaatc taccggattg caagcggtta aagaacgtta acctcgcgag
     1501 gaacaccttc catggacaag taccagagag tttcaagaac ttcgagagct tatcttactt
     1561 ctcgttatcg aattcgagtt tggctaatat ctcttcagcg cttgggatac ttcagcattg
     1621 caagaacttg acgactttgg ttcttacatt gaatttccat ggagaggctt tacccgatga
     1681 ttcaagtctt catttcgaga agcttaaggt gcttgtagtg gcgaattgta ggcttactgg
     1741 ttcgatgccg aggtggttaa gctcgagtaa tgaacttcag ttgttggatc tttcttggaa
     1801 ccgtttaacc ggcgctatcc cgagctggat tggtgacttc aaggctctgt tctacttgga
     1861 tttatctaac aactcgttta caggagagat ccctaagagc ttaactaagt tagagagtct
     1921 cactagccgt aatatctcag tcaatgagcc atctcctgat ttcccgttct ttatgaaaag
     1981 aaacgagagc gcgagagcgt tgcaatacaa tcagattttc gggttcccgc caacaattga
     2041 gcttggtcat aacaatctct ctggacctat ttgggaggag tttggtaatc tgaagaagct
     2101 tcatgtgttt gatttgaaat ggaatgcatt atctggatca atacctagct cgctttctgg
     2161 tatgacgagc ttggaagctc tcgatctctc taataaccgt ctttcgggtt cgatcccggt
     2221 ttctctgcaa cagctctcgt ttctgtcgaa gttcagtgtt gcttataaca atctctcggg
     2281 agtaatacct tccggtggtc agtttcagac gtttccaaac tcgagctttg agagtaacca
     2341 tctctgcggg gaacacagat tcccctgttc tgaaggtact gagagtgcat tgatcaaacg
     2401 gtcaagaaga agcctggagg tgctgttcca gggaccaggg agcgacaaaa ctcacacatg
     2461 cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct tccccccaaa
     2521 acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg tggtggacgt
     2581 gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa
     2641 tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg tcagcgtcct
     2701 caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg tctccaacaa
     2761 agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc cccgagaacc
     2821 acaggtgtac accctgcccc catcccggga ggagatgacc aagaaccagg tcagcctgac
     2881 ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga gcaatgggca
     2941 gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct ccttcttcct
     3001 ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct tctcatgctc
     3061 cgtgatgcat gagggtctgc acaaccacta cacgcagaag agcctctccc tgtctccggg
     3121 taaatctaga gggcccttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     3181 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     3241 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     3301 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3361 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3421 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3481 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga cctgcaggca
     3541 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3601 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3661 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3721 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3781 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3841 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3901 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3961 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4021 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4081 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4141 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4201 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4261 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4321 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4381 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4441 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4501 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4561 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4621 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4681 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4741 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4801 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4861 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4921 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4981 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5041 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5101 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5161 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5221 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5281 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5341 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5401 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5461 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5521 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5581 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5641 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5701 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5761 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5821 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5881 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     5941 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6001 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6121 attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
