LOCUS       sequence-298828-        6293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RGFR3 X052_pECIA2 should be used with prey vector RGFR3 
            X052_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6293)
  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 6293)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6293
                     /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             3275..3316
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3326..3343
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3366..3383)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3416..3550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3440..3459)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3494..3513
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3694..3710)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3694..3710)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3707..3729)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3718..3734
                     /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(3742..3748,3749..3766,3767..3772))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3787..3808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3925..3942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4096..4684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4176..4195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4855..5646,5647..5715))
                     /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     5478..5497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5716..5820)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5888..5906
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5944..5966)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6066..6085
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6279..6293,1..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 cagggaccag ggagcgacaa
     2581 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct
     2641 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
     2701 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
     2761 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
     2821 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
     2881 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
     2941 gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga ccaagaacca
     3001 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
     3061 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
     3121 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
     3181 cttctcatgc tccgtgatgc atgagggtct gcacaaccac tacacgcaga agagcctctc
     3241 cctgtctccg ggtaaatcta gagggccctt cgaaggtaag cctatcccta accctctcct
     3301 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     3361 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     3421 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3481 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3541 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3601 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     3661 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3721 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3781 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3841 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3901 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4501 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4921 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5041 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5281 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5521 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5701 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5761 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5821 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5881 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5941 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6001 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6061 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6121 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     6181 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6241 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gtt
//
