LOCUS       sequence-288213-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector EFR D5_pECIA14 should be used with bait vector EFR 
            D5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  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 7073)
  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: EFR D5_pECIA14
FEATURES             Location/Qualifiers
     source          1..7073
                     /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             2369..2392
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4067..4090
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4106..4123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4146..4163)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4196..4330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4220..4239)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4274..4293
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4474..4490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4474..4490)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4487..4509)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4498..4514
                     /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(4522..4528,4529..4546,4547..4552))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4567..4588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4705..4722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4876..5464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4956..4975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5635..6426,6427..6495))
                     /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     6258..6277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6496..6600)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6668..6686
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6724..6746)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6846..6865
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7059..7073,1..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 tctaatgaga ctgatatgca
      541 agctttgctt gagttcaagt ctcaagtttc tgaaaacaac aagagagagg tcttggcttc
      601 atggaatcac tcctccccat tttgtaattg gattggagtc acatgtggcc gcaggagaga
      661 aagagttata agtttgaacc ttggaggatt caagttaacc ggtgtgatct caccctccat
      721 tggtaatctc tcctttctta gattacttaa tcttgcagac aactcttttg gaagtaccat
      781 ccctcaaaag gtgggaaggc tatttaggct tcagtacttg aacatgagct ataatcttct
      841 cgaaggaagg attccgtcta gtctttctaa ctgctctaga ctgtcgaccg ttgatttatc
      901 gtcaaaccat cttggacatg gtgttccttc agaactaggt tcactttcta agcttgccat
      961 tctggatctt agcaaaaaca accttactgg aaattttcct gcatctttag gaaacttgac
     1021 gtcacttcag aagcttgact ttgcatataa ccagatgaga ggtgagattc cagacgaagt
     1081 agctagattg actcaaatgg tgtttttcca aatagcactg aatagttttt caggtggttt
     1141 tcctcctgca ttgtacaaca tctcctctct tgagtctcta tctctagctg acaatagctt
     1201 ttcgggtaat cttagggctg attttggtta tcttctacca aatctaagaa gacttctttt
     1261 gggaacaaat cagttcactg gagctattcc caaaacactt gccaatatct caagccttga
     1321 aaggtttgat atctcatcta attacctgtc tggtagtatc cctttgagct ttggaaagtt
     1381 acgtaatctg tggtggttag ggattcgtaa taactctctt ggaaataact cgtccagtgg
     1441 tcttgaattt attggggctg tggcgaactg cactcaatta gagtacttag atgttggtta
     1501 caatagactt ggaggtgagc ttcctgcttc tatagccaat ctgtccacta cattgactag
     1561 tctgttcctt ggacaaaatc ttatctctgg aaccattcct catgacatcg ggaatcttgt
     1621 aagcctgcaa gaactcagct tagaaacaaa tatgttgagt ggagaacttc ccgtctcttt
     1681 cgggaagctt ttgaacttgc aggttgtgga tctgtattca aatgcaatat cgggggaaat
     1741 accatcttat tttggcaaca tgactcggtt gcagaagctc catttgaata gcaatagttt
     1801 ccacggaaga atccctcaga gtcttggacg ttgtcgatac ttgctagacc tgtggatgga
     1861 tacaaatagg ttgaatggga ctatacctca ggaaatactg caaattccat ccctcgctta
     1921 catagatttg tcaaacaatt tcttgacagg ccattttcca gaagaagttg gaaagttaga
     1981 acttcttgtt ggactaggtg cttcgtacaa caaattatca ggaaagatgc cacaagctat
     2041 agggggttgt ctttcgatgg aatttctctt tatgcaagga aattcgtttg acggagccat
     2101 tccagatata agccggttgg taagcctaaa gaatgttgac ttctccaaca acaatctctc
     2161 tggccggata cctcgatatc tggccagtct tccttcgctg cgaaatctga atctttctat
     2221 gaacaagttt gagggaaggg tgccaacaac aggagtgttt cgaaatgcta cagcagtttc
     2281 tgtttttggt aacacaaata tttgcggagg cgtccgagaa atgcaactaa agccatgcat
     2341 tgtacaggca tctccaagga agagaaagct ggaggtgctg ttccagggac caagcggcaa
     2401 tcccggcggc gatctggccc cccagatgct gcgcgagctg caggagacca atgccgccct
     2461 gcaggacgtg cgcgagctgc tgcgccaccg cgtgaaggag atcaccttcc tgaagaacac
     2521 cgtgatggag tgcgacgcct gcggcatgca gccctctaga ggctccggct ccatcatccc
     2581 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     2641 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     2701 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     2761 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     2821 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     2881 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     2941 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     3001 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     3061 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     3121 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggagg
     3181 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     3241 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     3301 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     3361 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     3421 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     3481 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     3541 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     3601 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     3661 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     3721 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     3781 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     3841 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     3901 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     3961 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     4021 cgacgccgcg cacccgggtt ctagaggaag cacctccggc gataacgatt acaaggatga
     4081 tgatgataag gccagcggca ccggtcatca tcaccatcac cattgagttt aaacccgctg
     4141 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     4201 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4261 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4321 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4381 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     4441 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4501 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4561 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4621 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5761 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6601 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6661 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6721 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6781 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6841 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6901 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6961 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7021 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
