LOCUS       sequence-296422-        7250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RGFR3 X052_pECIA14 should be used with bait vector RGFR3
            X052_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7250)
  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 7250)
  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: RGFR3 X052_pECIA14
FEATURES             Location/Qualifiers
     source          1..7250
                     /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             2546..2569
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4244..4267
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4283..4300
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4323..4340)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4373..4507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4397..4416)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4451..4470
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4651..4667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4651..4667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4664..4686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4675..4691
                     /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(4699..4705,4706..4723,4724..4729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4744..4765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4882..4899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5053..5641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5133..5152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5812..6603,6604..6672))
                     /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     6435..6454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6673..6777)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6845..6863
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6901..6923)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7023..7042
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7236..7250,1..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 tctctcgacc aacaaggtca
      541 agctctcttg tcatggaagt ctcaactgaa catctccggc gacgcttttt cctcctggca
      601 cgtcgccgac acatctccct gcaattgggt cggcgtaaaa tgtaaccgta gaggtgaagt
      661 ttcggagata cagctcaaag gtatggactt gcaaggttct ctgccggtga ctagtctccg
      721 gagcctcaag tctcttactt ccctcacttt atcttcactg aatctcaccg gagtaatccc
      781 caaggagatc ggagacttca cggagcttga gttactcgat ttgtcggata attctctctc
      841 cggtgatatc cctgtggaaa tcttcaggct caagaaactc aagactctgt ctctgaacac
      901 taacaatcta gaaggtcata ttccgatgga gattgggaat ctttcgggtc tcgtagagct
      961 tatgcttttc gacaacaagc tatccggaga gatcccaagg agtatcggag agctcaagaa
     1021 tctacaagtc ttacgtgctg gtgggaacaa gaatctaaga ggtgagcttc cttgggagat
     1081 aggtaactgc gagaatcttg taatgcttgg tctcgctgag actagtctct ccgggaaact
     1141 tccggcgtcg attggaaacc tgaaacgtgt tcagacaata gcaatttaca cgtcactctt
     1201 gtctggtcca attccggatg agattggata ctgcacagag ctccaaaacc tctacttgta
     1261 tcagaattca atctcaggat cgatccctac caccattgga ggtttaaaga agctgcagag
     1321 tctactttta tggcagaaca atctcgttgg gaaaatccca accgagcttg ggaactgtcc
     1381 cgagctctgg cttatcgatt tttccgagaa tctcctcacc gggaccatcc caaggagctt
     1441 cggaaaactt gagaatctac aagagcttca gctgagtgta aaccagatct caggaacaat
     1501 ccctgaagag ctaacgaatt gcacgaagct gacgcattta gagatcgaca ataatctcat
     1561 cacaggagag attccgtcat tgatgagtaa cctaagaagc ttaaccatgt tcttcgcatg
     1621 gcagaacaag ttaactggaa acatccctca aagtctctcc cagtgtcgtg agcttcaagc
     1681 catcgatctc tcttacaaca gtctctctgg ctccatccca aaagagatat tcgggttacg
     1741 aaacctcacg aagcttcttc ttctctccaa tgatttgtcg ggtttcatac ctccggatat
     1801 tggaaactgt acgaatcttt accggttaag actcaacggt aacagactcg ccggaagtat
     1861 cccgtcggaa atcgggaatc tgaaaaatct caactttgtt gatataagtg agaaccgtct
     1921 cgtcgggtca atccctccgg cgatctctgg ttgtgaaagc ctcgagtttc tcgatcttca
     1981 cacgaatagt ctctccggtt cattgctcgg tactacactc ccaaagagct tgaagttcat
     2041 cgatttctcc gataatgctc tgtccagtac tctgcctccg ggaataggat tgttgacaga
     2101 gcttacaaag ctcaatctcg ccaagaacag attatccggc gaaatcccta gagaaatctc
     2161 cacttgtcgg agtctacagc ttctcaatct cggcgaaaat gacttttctg gtgaaatccc
     2221 tgacgaattg ggtcaaatcc cgtctctggc gatatctctt aatctcagct gcaataggtt
     2281 cgtcggagaa atcccgtcta gattctccga cttgaaaaac cttggagtgc tcgacgtatc
     2341 tcacaaccaa ctcaccggaa acttaaacgt tttaacggat ttgcaaaacc tcgtctctct
     2401 caacatctcc tacaacgatt tctccggcga tttacccaac acgccctttt tccggcgact
     2461 tccactttct gatctcgcct caaacagagg gctttacatc tcgaacgcaa tctcgacccg
     2521 acccgacccg acgaccagga acagcctgga ggtgctgttc cagggaccaa gcggcaatcc
     2581 cggcggcgat ctggcccccc agatgctgcg cgagctgcag gagaccaatg ccgccctgca
     2641 ggacgtgcgc gagctgctgc gccaccgcgt gaaggagatc accttcctga agaacaccgt
     2701 gatggagtgc gacgcctgcg gcatgcagcc ctctagaggc tccggctcca tcatcccagt
     2761 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     2821 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     2881 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     2941 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     3001 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     3061 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     3121 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     3181 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     3241 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     3301 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     3361 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     3421 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     3481 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     3541 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     3601 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     3661 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     3721 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     3781 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     3841 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     3901 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     3961 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     4021 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     4081 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     4141 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     4201 cgccgcgcac ccgggttcta gaggaagcac ctccggcgat aacgattaca aggatgatga
     4261 tgataaggcc agcggcaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     4321 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     4381 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4441 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4501 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4561 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     4621 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4681 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4741 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5221 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5461 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5881 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6781 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6841 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     6901 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6961 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7021 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     7081 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     7141 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     7201 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
