LOCUS       sequence-275966-        7649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector EMS1 X044_pECIA14 should be used with bait vector EMS1 
            X044_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7649)
  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 7649)
  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: EMS1 X044_pECIA14
FEATURES             Location/Qualifiers
     source          1..7649
                     /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             2945..2968
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4643..4666
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4682..4699
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4722..4739)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4772..4906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4796..4815)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4850..4869
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5050..5066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5050..5066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5063..5085)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5074..5090
                     /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(5098..5104,5105..5122,5123..5128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5143..5164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5281..5298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5452..6040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5532..5551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6211..7002,7003..7071))
                     /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     6834..6853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7072..7176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7244..7262
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7300..7322)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7422..7441
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7635..7649,1..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 tccgccattg ttgatcttag
      541 ctcggaaaca acttctctca tctccttcaa gcgctcacta gaaaacccat ctctcctttc
      601 ttcatggaac gtttcgtctt ccgcgtctca ctgcgattgg gttggcgtta cgtgtctgct
      661 tggacgagtc aactcgctct ccttaccgtc tttgtctctg agaggccaga ttccgaagga
      721 aatttcgagt ttgaagaacc ttagagagct ttgtcttgcc ggaaatcaat tctccgggaa
      781 gattccgcct gagatttgga atctaaaaca tctccaaacc cttgacctct caggaaattc
      841 tctcaccgga cttctcccga ggctactcag tgagttacct cagcttctct acctcgacct
      901 tagtgacaac cacttctccg gttcacttcc tccatcgttc ttcatcagct tacctgcact
      961 gtcttctctt gacgtctcca acaactctct ctccggcgaa atccctcctg aaatcgggaa
     1021 actgagtaac ctctccaatc tttacatggg actcaactca ttttccggtc aaattccgtc
     1081 ggagattggt aacatctcgt tgctgaagaa ctttgctgca ccttcttgct tcttcaatgg
     1141 accgttacca aaggagattt cgaaactgaa acacttagca aagctcgacc tttcttataa
     1201 tccactcaag tgttccattc ccaaatcgtt tggagagttg cataatctga gtatactaaa
     1261 tcttgtctcc gcagagctta ttggattgat cccaccagag cttggtaatt gtaaaagcct
     1321 caagtcactg atgctgtctt tcaattctct ctctggacct ttgccattgg agctttcaga
     1381 gattcctctg ttgacgtttt ctgctgaaag gaatcagctt tcgggttcat tgccttcatg
     1441 gatggggaaa tggaaggtac tggactcact tctgttagct aacaatcggt tttcaggtga
     1501 aatccctcat gagattgagg attgtccaat gttgaaacat ctcagtctag ctagcaactt
     1561 gttaagtggt tcgattccta gggagctctg tggttcaggt tcgttggagg cgattgatct
     1621 ctctgggaat ttgctttctg gaacaattga ggaggtcttt gatggatgtt cctctcttgg
     1681 agagttgtta cttactaata atcagatcaa tggctctatt cctgaggatc tttggaagct
     1741 tccattgatg gcacttgact tggattccaa caacttcaca ggagaaatcc caaagagtct
     1801 ctggaaatca acgaacttga tggagtttac agcttcttat aatcgtctag aaggttattt
     1861 gcctgcagag attggtaatg cagcttcact gaagcggttg gttctcagtg ataatcagtt
     1921 aacaggtgaa atcccaagag agattggaaa actcacatct ttgtctgtct tgaatctgaa
     1981 tgcaaacatg tttcaaggga agataccagt tgagctcggt gattgtactt ctttgactac
     2041 attggatcta ggcagcaaca atcttcaagg acagattcca gacaaaatca ctgctctcgc
     2101 tcagcttcag tgcctggttc tctcttacaa caatctctcc gggtcaattc catcaaaacc
     2161 ctctgcttat ttccatcaga ttgagatgcc tgatctcagc tttcttcagc accatgggat
     2221 atttgatctc tcctacaata gattgtctgg ccctatacca gaagaacttg gcgagtgttt
     2281 ggtattggta gagatttcac tgagcaacaa ccatctctct ggagagattc cagcttcact
     2341 ctctcgtttg accaacctca caatcctgga tctatctgga aatgcactga ctggatctat
     2401 tccaaaggag atgggcaact ctcttaagtt gcagggatta aacttggcga ataatcagct
     2461 caatggccat attccagaaa gctttggctt gttaggtagc ttggtgaagc taaacttgac
     2521 taagaacaag cttgatggac cggttcctgc ttctcttggg aacttgaaag agcttacaca
     2581 catggacttg agctttaaca acctaagtgg cgagctttca tcagaactgt ctacaatgga
     2641 aaaacttgtc ggcctctaca ttgagcagaa taagttcacc ggggaaatac cttcagaact
     2701 tggtaatctt acgcagctgg agtaccttga tgtttctgag aacttgttat ctggagagat
     2761 accaacaaag atctgtggat tgcccaatct ggagttcttg aatctagcca agaacaatct
     2821 gcgaggagag gtgccgagtg atggtgtttg tcaggatcct tccaaggcat tgctttcagg
     2881 gaacaaagag ttgtgtggca gagtggttgg atcagattgc aagatcgagg gaactaagtt
     2941 gaggctggag gtgctgttcc agggaccaag cggcaatccc ggcggcgatc tggcccccca
     3001 gatgctgcgc gagctgcagg agaccaatgc cgccctgcag gacgtgcgcg agctgctgcg
     3061 ccaccgcgtg aaggagatca ccttcctgaa gaacaccgtg atggagtgcg acgcctgcgg
     3121 catgcagccc tctagaggct ccggctccat catcccagtt gaggaggaga acccggactt
     3181 ctggaaccgc gaggcagccg aggccctggg tgccgccaag aagctgcagc ctgcacagac
     3241 agccgccaag aacctcatca tcttcctggg cgatgggatg ggggtgtcta cggtgacagc
     3301 tgccaggatc ctaaaagggc agaagaagga caaactgggg cctgagatac ccctggccat
     3361 ggaccgcttc ccatatgtgg ctctgtccaa gacatacaat gtagacaaac atgtgccaga
     3421 cagtggagcc acagccacgg cctacctgtg cggggtcaag ggcaacttcc agaccattgg
     3481 cttgagtgca gccgcccgct ttaaccagtg caacacgaca cgcggcaacg aggtcatctc
     3541 cgtgatgaat cgggccaaga aagcagggaa gtcagtggga gtggtaacca ccacacgagt
     3601 gcagcacgcc tcgccagccg gcacctacgc ccacacggtg aaccgcaact ggtactcgga
     3661 cgccgacgtg cctgcctcgg cccgccagga ggggtgccag gacatcgcta cgcagctcat
     3721 ctccaacatg gacattgacg tgatcctagg tggaggccga aagtacatgt ttcgcatggg
     3781 aaccccagac cctgagtacc cagatgacta cagccaaggt gggaccaggc tggacgggaa
     3841 gaatctggtg caggaatggc tggcgaagcg ccagggtgcc cggtatgtgt ggaaccgcac
     3901 tgagctcatg caggcttccc tggacccgtc tgtgacccat ctcatgggtc tctttgagcc
     3961 tggagacatg aaatacgaga tccaccgaga ctccacactg gacccctccc tgatggagat
     4021 gacagaggct gccctgcgcc tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga
     4081 gggtggtcgc atcgaccatg gtcatcatga aagcagggct taccgggcac tgactgagac
     4141 gatcatgttc gacgacgcca ttgagagggc gggccagctc accagcgagg aggacacgct
     4201 gagcctcgtc actgccgacc actcccacgt cttctccttc ggaggctacc ccctgcgagg
     4261 gagctccatc ttcgggctgg cccctggcaa ggcccgggac aggaaggcct acacggtcct
     4321 cctatacgga aacggtccag gctatgtgct caaggacggc gcccggccgg atgttaccga
     4381 gagcgagagc gggagccccg agtatcggca gcagtcagca gtgcccctgg acgaagagac
     4441 ccacgcaggc gaggacgtgg cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg
     4501 cgtgcaggag cagaccttca tagcgcacgt catggccttc gccgcctgcc tggagcccta
     4561 caccgcctgc gacctggcgc cccccgccgg caccaccgac gccgcgcacc cgggttctag
     4621 aggaagcacc tccggcgata acgattacaa ggatgatgat gataaggcca gcggcaccgg
     4681 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     4741 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     4801 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4861 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4921 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4981 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc
     5041 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5101 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5161 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5221 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5281 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5341 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5641 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5881 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6001 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6121 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6241 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6301 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6361 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6421 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6481 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6541 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6601 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6661 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6721 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6781 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6841 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6901 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6961 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7021 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7081 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7141 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7201 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7261 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7321 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7381 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7441 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7501 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7561 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7621 aacgccaggg ttttcccagt cacgacgtt
//
