LOCUS       sequence-296643-        5711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RHS16 N12_pECIA2 should be used with prey vector RHS16 
            N12_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5711)
  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 5711)
  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: RHS16 N12_pECIA2
FEATURES             Location/Qualifiers
     source          1..5711
                     /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             1964..1987
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2693..2734
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2744..2761
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2784..2801)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2834..2968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2858..2877)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2912..2931
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3112..3128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3112..3128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3125..3147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3136..3152
                     /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(3160..3166,3167..3184,3185..3190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3205..3226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3343..3360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3514..4102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3594..3613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4273..5064,5065..5133))
                     /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     4896..4915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5134..5238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5306..5324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5362..5384)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5484..5503
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5697..5711,1..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 caagcaggtt tcatcagcat
      541 agattgtggg tcacccccta atattaacta tgtagacact gatactggta tcagttacac
      601 ctgggatgca cctttcatca acgctggagt aaatctgaac gtctctgaag aatacggtta
      661 cccgaaaaac ccggttttgc ctttcccgct tgctgatgta agatcttttc ctcaaggaaa
      721 caggaactgt tacaccttaa ccccctccga tggaaaaggg aatctttatc tcatcagagc
      781 ttcgtttatg tatggaaact acgatggtaa aaatgcttta cctgagtttg atctctacgt
      841 aaacgtgaat ttctggacct cggtgaaact cagaaatgct tctgagaacg tcatcaagga
      901 gatcctcagc tttgcagagt cagatacaat atatgtttgc cttgtgaaca aaggtaaagg
      961 aactcctttt atttcagcgt tggagcttcg accaatgaac agctccatct atggaactga
     1021 gtttggaaga aatgtctcat tggtactcta tcaaagatgg gatacaggat atttgaatgg
     1081 aacaggacgg taccaaaagg atacatatga tcgtatttgg tctccgtact ctccagtgtc
     1141 ttggaataca actatgacta ctgggtatat cgatattttt cagtctggtt ataggccacc
     1201 agatgaagtt attaagacag ctgcttcgcc taaaagtgac gatgaaccat tggaactgtc
     1261 ttggacatca agcgatccag atactcggtt ttatgcgtac ctttattttg cagaacttga
     1321 aaatctcaag aggaatgaga gcagagaaat caagatattc tggaatggat cgcctgtttc
     1381 aggagctttt aacccctctc ctgagtattc aatgacagtg tctaactcac gagctttcac
     1441 tggtaaagat cactggattt ctgttcagaa gaccgcagag tcaacgcgtc caccaatact
     1501 caatgctatt gagatttttt cagcacaatc tctggatgaa ttttacacca gaattgacga
     1561 tgttcaagcc atagaaagca taaaatcaac gtataaagtg aataagattt ggactggtga
     1621 tccttgctct cccagactct tcccttggga aggtattgga tgcagctaca acacttctag
     1681 ttatcaaatc aagtccctaa atcttagttc aagtggatta catggaccaa tagcgttcgc
     1741 atttagaaat ctctcccttc ttgagtcatt ggatttatca aacaacaacc tcaaaggaat
     1801 tgtgccggaa tttttggctg atctaaaata tctgaagtcc ctgaatttga aaggaaataa
     1861 cttgactggg tttatcccaa gatctcttag gaaaagagca acggctaatg gacttgcact
     1921 cagtgtcgat gaacaaaaca tttgtcactc acgttcatgt cggctggagg tgctgttcca
     1981 gggaccaggg agcgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg
     2041 gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac
     2101 ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa
     2161 ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta
     2221 caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg
     2281 caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
     2341 ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga
     2401 ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga
     2461 catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc
     2521 cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag
     2581 gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gagggtctgc acaaccacta
     2641 cacgcagaag agcctctccc tgtctccggg taaatctaga gggcccttcg aaggtaagcc
     2701 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     2761 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     2821 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     2881 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2941 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3001 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3061 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     3121 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3181 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3241 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3301 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3361 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3421 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3481 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3541 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3961 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4021 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4081 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4141 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4201 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4261 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4321 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4381 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4441 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4501 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4561 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4621 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4681 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4741 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4801 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4861 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4921 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4981 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5041 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5101 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5161 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5221 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5281 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     5341 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     5401 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     5461 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     5521 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     5581 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5641 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5701 gtcacgacgt t
//
