LOCUS       sequence-273962-        6146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector MDIS1 P4_pECIA14 should be used with bait vector MDIS1 
            P4_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6146)
  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 6146)
  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: MDIS1 P4_pECIA14
FEATURES             Location/Qualifiers
     source          1..6146
                     /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             1442..1465
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3140..3163
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3179..3196
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3269..3403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3293..3312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3347..3366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3547..3563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3547..3563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3560..3582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3571..3587
                     /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(3595..3601,3602..3619,3620..3625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3640..3661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3778..3795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3949..4537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4029..4048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4708..5499,5500..5568))
                     /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     5331..5350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5569..5673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5741..5759
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5797..5819)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5919..5938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6132..6146,1..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 tccctcgaat ccgaaggatt
      541 tgtattgttg aaattccggg caagagttga ttctgatcct catggaactc ttgcaaattg
      601 gaatgtttct gatcatgatc atttctgttc ttggtttggt gttacgtgtg tcgacaataa
      661 agtgcagatg ctgaatctta gtggttgttc tttgggagga actttagctc cggagcttag
      721 tcaattgagt gaattaagat ctctaatact atccaagaac aaactctctg gtgacattcc
      781 aaacgaattt gcgagtttcg cgaaattaga gttcttggat ttgcgagata ataacttaaa
      841 cggagtagtt ccacccgagc taaataaagt gttgacacca gaaaacttgt tgctttctgg
      901 taacaaattt gcgggtttta tgaccgtaaa gttcctgaga cttcaatcgc tgtataaagt
      961 ccagatgaac aagaatcgag agctatcttc ggtttccgct gatgtcctcg actgtgtcaa
     1021 tagaaaactt ggatactgtg tttcaaggag aagtttaata acaagaaaca aggcaaaggc
     1081 attcgtattg cggattagag caacttcaag acattacatg gtgcgaagag aatctcatgg
     1141 gaagaattat gtagtgaatt atcacccaag tgagaacgaa actagtatct tcaaaagacg
     1201 tgagttactc gaggaaacaa gcaatttagc ggctatgcct gcgcctgata cgcccagtcc
     1261 ttctcctgag attataacta tagtgtttcc tcgaagcagc gggtcgtttc cagcattaac
     1321 taatgcaaag aagagaatac ctccattgat ccctccttct tctcctcctc ctctacctac
     1381 caacaacacc atcgctagtg atccaccgag gaaattcgaa gaaaaatcga aagggtttaa
     1441 gctggaggtg ctgttccagg gaccaagcgg caatcccggc ggcgatctgg ccccccagat
     1501 gctgcgcgag ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc tgctgcgcca
     1561 ccgcgtgaag gagatcacct tcctgaagaa caccgtgatg gagtgcgacg cctgcggcat
     1621 gcagccctct agaggctccg gctccatcat cccagttgag gaggagaacc cggacttctg
     1681 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     1741 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     1801 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     1861 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     1921 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     1981 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     2041 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     2101 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     2161 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     2221 caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac
     2281 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     2341 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     2401 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     2461 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     2521 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     2581 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     2641 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     2701 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     2761 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     2821 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     2881 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     2941 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     3001 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     3061 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gttctagagg
     3121 aagcacctcc ggcgataacg attacaagga tgatgatgat aaggccagcg gcaccggtca
     3181 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     3241 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3301 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3361 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3421 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3481 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     3541 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3601 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3661 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3721 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3781 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3841 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4381 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4741 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4801 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4861 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4921 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4981 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5041 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5101 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5161 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5221 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5281 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5341 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5401 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5461 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5521 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5581 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5641 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5701 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5761 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5821 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5881 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5941 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6001 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6061 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6121 gccagggttt tcccagtcac gacgtt
//
