LOCUS       sequence-297005-        5147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector MDIS2/MRH1 P1_pECIA2 should be used with prey vector 
            MDIS2/MRH1 P1_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5147)
  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 5147)
  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: MDIS2/MRH1 P1_pECIA2
FEATURES             Location/Qualifiers
     source          1..5147
                     /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             1400..1423
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2129..2170
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2180..2197
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2220..2237)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2270..2404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2294..2313)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2348..2367
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2548..2564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2548..2564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2561..2583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2572..2588
                     /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(2596..2602,2603..2620,2621..2626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2641..2662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2779..2796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2950..3538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3030..3049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3709..4500,4501..4569))
                     /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     4332..4351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4570..4674)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4742..4760
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4798..4820)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4920..4939
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5133..5147,1..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 tccctcactt cccaaggatc
      541 cgcattgttg aagttccggg caagagtcaa ctcagatcct catgggactc ttgcgaattg
      601 gaatgtttct ggtattaatg atctctgcta ttggtcaggg gttacttgtg ttgacggtaa
      661 agtgcagatt ctggatctta gcggatattc tttggaagga acgttagctc ctgagcttag
      721 ccaattgagt gacttaagat ctctaatact ctctcggaac catttctccg gtggaattcc
      781 aaaggagtat gggagtttcg aaaacttgga agtcctggat ttgagagaga atgatttgag
      841 cggacaaatt ccacctgagc taagcaatgg cttatcgcta aaacacttgt tgctttctgg
      901 taacaaattt tcggatgata tgagaataaa gatcgtgaga cttcaatctt cgtatgaagt
      961 tcggttaaag aagagtccaa agttatcacc tttggctgtt ctcggctgta ttaacagaaa
     1021 acttggacac tgtgtttcaa gaaaccgtat cattcaagta aaaaaggtag aagctattgt
     1081 attccggatc aaagcaacct ccagacgttt cttgaaagca ttccctagct ttttggagga
     1141 aactgatatc tacaaacgac gtgagctact cgaggagaca agtaatttag cggctgagcc
     1201 tgctccttca gctcctagtc cttctcccgg gattataact gaagcttcac ctcgaagcag
     1261 cgggtctttc cctgcagtaa ccaatgcaaa gaagagaaga ccacctttag tccctcctgt
     1321 accttctcct gataagggca gtactagtcc tgatatctct aagaatcagc cacaagataa
     1381 taaacagtca aaaggatcac tggaggtgct gttccaggga ccagggagcg acaaaactca
     1441 cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc
     1501 cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt
     1561 ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt
     1621 gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag
     1681 cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc
     1741 caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg
     1801 agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag
     1861 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     1921 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     1981 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     2041 atgctccgtg atgcatgagg gtctgcacaa ccactacacg cagaagagcc tctccctgtc
     2101 tccgggtaaa tctagagggc ccttcgaagg taagcctatc cctaaccctc tcctcggtct
     2161 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     2221 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     2281 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2341 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2401 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2461 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     2521 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2581 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2641 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2701 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2761 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2821 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2881 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2941 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3001 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3061 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3121 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3181 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3241 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3301 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3361 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3421 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3481 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3661 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3721 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3781 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3841 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3901 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3961 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4021 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4081 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4141 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4201 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4261 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4321 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4381 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4441 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4501 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4561 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4621 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4681 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4741 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4801 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4861 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4921 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     4981 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     5041 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5101 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
