LOCUS       sequence-298838-        6320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RGFR2 X055_pECIA2 should be used with prey vector RGFR2 
            X055_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6320)
  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 6320)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6320
                     /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             3302..3343
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3353..3370
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3393..3410)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3443..3577
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3467..3486)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3521..3540
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3721..3737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3721..3737)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3734..3756)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3745..3761
                     /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(3769..3775,3776..3793,3794..3799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3814..3835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3952..3969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4123..4711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4203..4222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4882..5673,5674..5742))
                     /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     5505..5524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5743..5847)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5915..5933
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5971..5993)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6093..6112
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6306..6320,1..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 ggaccaggga gcgacaaaac tcacacatgc ccaccgtgcc cagcacctga
     2641 actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat
     2701 ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt
     2761 caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga
     2821 ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg
     2881 gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga
     2941 gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc
     3001 atcccgggag gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta
     3061 tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac
     3121 cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga
     3181 caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg agggtctgca
     3241 caaccactac acgcagaaga gcctctccct gtctccgggt aaatctagag ggcccttcga
     3301 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
     3361 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     3421 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3481 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3541 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3601 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3661 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4561 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4801 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5041 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5401 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5761 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5821 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5881 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5941 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6001 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6061 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6121 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6181 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6241 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6301 gttttcccag tcacgacgtt
//
