LOCUS       sequence-298818-        6308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RGFR1 X077_pECIA2 should be used with prey vector RGFR1 
            X077_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6308)
  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 6308)
  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: RGFR1 X077_pECIA2
FEATURES             Location/Qualifiers
     source          1..6308
                     /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             2561..2584
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3290..3331
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3341..3358
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3381..3398)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3431..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3455..3474)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3509..3528
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3709..3725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3709..3725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3722..3744)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3733..3749
                     /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(3757..3763,3764..3781,3782..3787))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3802..3823
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3940..3957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4111..4699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4191..4210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4870..5661,5662..5730))
                     /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     5493..5512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5731..5835)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5903..5921
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5959..5981)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6081..6100
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6294..6308,1..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 tctatccttt actcatggct
      541 tcactcttct tctcctacac cttcttctct ttctctcttc aactggaact ctatagacaa
      601 cactccttgc aacaactgga catttatcac atgttcttct caaggtttca tcaccgacat
      661 agacattgaa tctgtccctc ttcagctctc cttaccgaag aatctcccag cgtttcgttc
      721 tctccagaaa ctcactatct ccggggccaa tctcaccgga actttaccgg aaagtcttgg
      781 tgactgcctc ggcctcaagg ttcttgatct tagctccaac ggcttagttg gtgacattcc
      841 ttggagttta agcaagctac gtaacctaga aacgctaatc ctcaactcaa accaactcac
      901 cggaaaaatc ccaccggata tcagcaaatg ttcgaaactc aagagcctga ttctcttcga
      961 caatcttctc accggaagta tccccacgga gctagggaag ctctctggtc tcgaagtaat
     1021 aagaatagga ggaaacaaag aaatctccgg ccaaatccca tcagagatcg gagattgttc
     1081 caatctaaca gttcttggtt tagccgagac aagcgtctca ggaaacttgc cttcttcctt
     1141 aggtaagctc aagaagctag aaacactctc gatatacaca acaatgataa gtggtgagat
     1201 tccttctgat ttagggaatt gctctgagct tgttgatctc ttcttgtacg agaatagctt
     1261 gtctggttcc atccctcgag aaatcggaca gctaacgaaa ctggaacagc tgtttctatg
     1321 gcagaacagt ctcgttggtg gtatccctga agaaatagga aactgcagca atctcaaaat
     1381 gatagatttg tctttaaacc tcctctctgg ttcgattcca agctccattg gtcgtttgtc
     1441 ttttcttgaa gagtttatga tcagtgacaa caagttctcc ggttcaatac caacaacaat
     1501 ctcaaactgc tcgagtcttg tccagttaca gctcgataag aatcagattt caggtctaat
     1561 accgtccgag cttggaacac tcacaaagct tactctgttc tttgcttggt ctaatcagct
     1621 agaaggaagt atccctcctg gtttagcaga ttgtactgat cttcaagcat tggatttgtc
     1681 gcgcaattcg ctaaccggaa caatcccttc cggtttgttt atgctaagga acctcacaaa
     1741 gcttctcttg atctcaaact ctctgtctgg ttttatacct caagagattg gtaattgcag
     1801 ctctttggtg agattgaggt taggtttcaa caggatcaca ggagagattc cttcgggtat
     1861 tggttctctt aagaagataa actttctcga tttctcgagt aataggctac atggaaaggt
     1921 tcctgatgag attggaagct gttcagagtt acagatgatt gatctcagta ataactctct
     1981 tgaaggttct ttgcctaatc cggtttcttc actctccggt ttacaagtac ttgatgtttc
     2041 agctaaccag ttttcaggca agattccagc gagtttagga cggcttgtgt cactgaacaa
     2101 actaatcttg agcaagaacc tgttctcagg atcgatacct acttctctag ggatgtgttc
     2161 agggcttcag cttcttgatc ttggaagcaa cgagctctct ggtgagattc cttcagagct
     2221 tggagatatc gagaatcttg aaatcgcgtt aaacttgagt agcaatagac tcactgggaa
     2281 gattccttca aagattgcgt ctctcaacaa gctctcgatt cttgatctct ctcacaacat
     2341 gcttgaagga gatcttgctc cactcgctaa tattgagaat ctagtctctc ttaacatatc
     2401 ttacaatagc ttctctggtt atcttccaga caacaagctt ttcagacaat tatctcctca
     2461 agatcttgaa ggaaacaaga aactctgttc ttcgactcaa gattcttgct ttctcaccta
     2521 cagaaagggt aatggacttg gagatgatgg tgatgcttct ctggaggtgc tgttccaggg
     2581 accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     2641 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     2701 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     2761 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     2821 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     2881 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     2941 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     3001 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     3061 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     3121 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     3181 gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca accactacac
     3241 gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag gtaagcctat
     3301 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     3361 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     3421 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3481 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3541 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3601 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3661 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3721 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3781 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3841 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3901 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5881 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5941 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6001 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6061 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6121 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6181 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6241 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6301 acgacgtt
//
