LOCUS       sequence-296428-        7277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RGFR2 X055_pECIA14 should be used with bait vector RGFR2
            X055_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  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 7277)
  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: RGFR2 X055_pECIA14
FEATURES             Location/Qualifiers
     source          1..7277
                     /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             2573..2596
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4271..4294
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4310..4327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4350..4367)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4400..4534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4424..4443)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4478..4497
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4678..4694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4678..4694)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4691..4713)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4702..4718
                     /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(4726..4732,4733..4750,4751..4756))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4771..4792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4909..4926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5160..5179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5839..6630,6631..6699))
                     /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     6462..6481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6700..6804)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6872..6890
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6928..6950)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7050..7069
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7263..7277,1..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 tcaaccaatg aagtctcagc
      541 tttgatttct tggcttcaca gctcgaactc gccgccaccg tcagtcttct ccggctggaa
      601 tccttctgat tctgacccat gtcaatggcc ttacattact tgttcctcct cagacaacaa
      661 actcgttaca gagatcaatg tcgtctctgt tcagttagct ttacctttcc ctcctaacat
      721 ttcttcgttt acttcacttc agaaactcgt tatctccaac actaatctca ccggagctat
      781 atcttctgag atcggagatt gttcggagct tatagttatc gatttaagct caaatagtct
      841 tgttggtgaa ataccttcga gtctagggaa gctcaagaat cttcaagaac tttgtttaaa
      901 ctccaatggc ctcacaggaa agatcccacc ggaactcggc gactgcgtta gcctcaagaa
      961 tcttgagatt ttcgataact acttatccga gaatctcccg ttggagctcg gaaagatctc
     1021 gactcttgag agcataagag ccggaggaaa ctcagagctt tcagggaaga tcccggagga
     1081 gatcggaaac tgtaggaatc tcaaggtctt agggttagca gcaacgaaaa tatccggttc
     1141 tttacctgtt tcgttgggtc aactaagcaa gctccagtct ttatctgtct attctacaat
     1201 gctctcaggt gagatcccta aagagcttgg aaactgctct gaacttatca atctgtttct
     1261 atacgacaat gacttgtccg gcacgcttcc aaaagaacta ggaaagcttc aaaacctaga
     1321 gaagatgctt ctatggcaga ataatctcca cggacctatc cctgaggaga ttggattcat
     1381 gaaaagctta aacgccattg atctctctat gaactatttc tcaggaacca tccctaaatc
     1441 gttcggtaac ttgtcgaatc tccaagagct tatgcttagc agcaacaaca tcaccgggtc
     1501 aataccttcg attctaagta actgcacaaa gcttgttcag ttccagattg acgccaatca
     1561 gatttcaggt ttgattccac cagagattgg attgcttaag gagctcaaca tcttcttggg
     1621 atggcagaat aagctagaag ggaatattcc ggacgagtta gctggttgtc agaatcttca
     1681 agctcttgac ttgtcacaga attatctcac cggatcatta cctgcaggtt tgtttcagct
     1741 tcgtaatctg actaagctct tgcttatatc taatgccatc tccggtgtta tcccactgga
     1801 gatcgggaac tgcacttcgc ttgtcagatt aaggcttgtc aataacagaa tcaccggaga
     1861 aatccctaaa ggtattggat ttcttcagaa ccttagcttc ctcgacttgt ctgagaacaa
     1921 tctctccggt ccagttcctt tggagataag taattgtaga caactccaga tgcttaactt
     1981 gagcaacaac actcttcaag gttatcttcc tctgtcttta tcatcgttaa caaagcttca
     2041 agtacttgat gtctcttcca atgacttgac tggcaagata cctgatagtc taggtcatct
     2101 catttcactc aaccggctca ttctaagtaa gaattctttc aatggagaaa tcccttcgtc
     2161 tctcggtcac tgcacgaatc ttcagcttct tgatctcagc agtaataaca tctctggaac
     2221 tataccggag gagctttttg acatccaaga tctagacatt gcgttaaact tgagctggaa
     2281 ttcattagat ggctttatcc cggaaaggat ttccgcgctt aaccgattat ccgtgctcga
     2341 tatttcgcat aacatgcttt caggcgacct ttctgcgctg tccggtctag aaaacttggt
     2401 ttctctgaac atctctcata acagattctc aggttatctt ccagatagta aagtgtttag
     2461 acagctgata ggagcagaga tggaaggaaa caatggactc tgttccaaag gtttcaggtc
     2521 ttgctttgta agtaacagtt cacagttaac tacacagcgt ggtgtgcact cactggaggt
     2581 gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     2641 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     2701 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca tgcagccctc
     2761 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2821 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2881 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2941 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     3001 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     3061 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     3121 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     3181 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     3241 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     3301 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     3361 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     3421 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     3481 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     3541 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     3601 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     3661 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     3721 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3781 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3841 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3901 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3961 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     4021 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     4081 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     4141 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     4201 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag gaagcacctc
     4261 cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc atcatcacca
     4321 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     4381 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     4441 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4501 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4561 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4621 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     4681 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4741 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4801 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4861 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6841 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6901 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6961 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     7021 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     7081 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     7141 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     7201 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7261 ttcccagtca cgacgtt
//
