LOCUS       sequence-270163-        6668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RHS16 N12_pECIA14 should be used with bait vector RHS16 
            N12_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6668)
  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 6668)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..6668
                     /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             3662..3685
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3701..3718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3741..3758)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3791..3925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3815..3834)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3869..3888
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4069..4085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4069..4085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4082..4104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4093..4109
                     /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(4117..4123,4124..4141,4142..4147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4162..4183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4300..4317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4471..5059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4551..4570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5230..6021,6022..6090))
                     /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     5853..5872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6091..6195)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6263..6281
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6319..6341)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6441..6460
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6654..6668,1..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 tggaatgggt 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 gggaccaagc ggcaatcccg gcggcgatct ggccccccag atgctgcgcg agctgcagga
     2041 gaccaatgcc gccctgcagg acgtgcgcga gctgctgcgc caccgcgtga aggagatcac
     2101 cttcctgaag aacaccgtga tggagtgcga cgcctgcggc atgcagccct ctagaggctc
     2161 cggctccatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     2221 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     2281 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     2341 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     2401 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     2461 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     2521 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     2581 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     2641 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     2701 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     2761 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     2821 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     2881 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     2941 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     3001 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     3061 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     3121 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     3181 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     3241 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     3301 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     3361 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     3421 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     3481 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     3541 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     3601 ccccgccggc accaccgacg ccgcgcaccc gggttctaga ggaagcacct ccggcgataa
     3661 cgattacaag gatgatgatg ataaggccag cggcaccggt catcatcacc atcaccattg
     3721 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     3781 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3841 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3901 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3961 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     4021 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     4081 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4141 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4261 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5341 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6241 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6301 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6361 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6421 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6481 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6541 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6601 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6661 acgacgtt
//
