LOCUS       sequence-275978-        7247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector RGI5 X047_pECIA14 should be used with bait vector RGI5 
            X047_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7247)
  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 7247)
  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: RGI5 X047_pECIA14
FEATURES             Location/Qualifiers
     source          1..7247
                     /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             2543..2566
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4241..4264
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4280..4297
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4320..4337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4370..4504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4394..4413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4448..4467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4648..4664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4648..4664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4661..4683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4672..4688
                     /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(4696..4702,4703..4720,4721..4726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4741..4762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4879..4896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5050..5638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5130..5149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5809..6600,6601..6669))
                     /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     6432..6451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6670..6774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6842..6860
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6898..6920)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7020..7039
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7233..7247,1..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 tcatcagatg gccaagctct
      541 tctctctctc aagagaccat caccatcact cttctcttct tgggacccac aagaccagac
      601 accatgttca tggtatggca ttacatgttc tgcagacaac agagtcatct ctgtctctat
      661 ccccgacact ttccttaacc tgtcttcaat ccctgacctc tcttctctct cctctctaca
      721 gttcctaaac ctctcctcca ccaacctctc tggcccaatc cctccttcat ttggaaaact
      781 tacccaccta aggcttctcg acctttcttc aaactctctc tctggtccaa tcccatcaga
      841 gcttggtcgc ctctctactc tccagttcct tatattgaac gccaacaagc tctcaggttc
      901 aatcccttca cagatttcga atctatttgc cttgcaagtg ctttgtcttc aagacaatct
      961 gttgaatggt tcgattcctt caagttttgg ctctttggtc tctcttcagc agttcagact
     1021 tggtggtaac actaatctcg gaggtcctat ccctgctcag ctagggtttc tgaaaaattt
     1081 gacaactcta gggtttgcag caagtggttt gtcaggttca atcccttcca catttgggaa
     1141 tttggtaaat ttgcaaactt tggctctgta tgacactgag atctctggta ccattcctcc
     1201 tcaactgggt ttgtgctcag agctcaggaa cttgtatttg cacatgaaca agctcactgg
     1261 atcaatccca aaggaactgg gtaagctcca gaagatcact agcttgcttc tatgggggaa
     1321 ttctttatct ggggtcatcc caccagagat ttccaactgt tcttccctcg tggtgtttga
     1381 tgtttcagct aacgatctaa ctggcgatat tcctggagat ttggggaagc ttgtttggtt
     1441 ggagcagctt cagttgtctg ataatatgtt taccggtcag attccttggg agttaagcaa
     1501 ctgttccagc ctcattgctc ttcagctgga caagaacaag ttatcaggct ccataccctc
     1561 acaaattgga aacctcaagt ctctgcagag ctttttcttg tgggagaact caatttcagg
     1621 aacaattccg tcttcttttg gcaactgcac agacctagtg gccctggacc tctctaggaa
     1681 caagctaaca ggaagaatac cagaagagct cttcagttta aagaggctca gcaaacttct
     1741 ccttctaggg aactccttat ccggtggatt accaaagagc gtagcaaaat gccagtcgct
     1801 tgtgaggcta agagtcggag agaaccaact ctcaggtcaa atccctaaag agataggcga
     1861 gttacagaac ctggtgtttc ttgatctcta catgaaccat ttctccggtg ggcttcctta
     1921 cgagatctct aacatcactg tccttgagct cttagatgtc cacaacaact acatcaccgg
     1981 agatattcct gctcagttgg ggaaccttgt gaatctagag cagcttgatt tgagcagaaa
     2041 cagcttcaca ggaaacatac cgctgagttt tgggaaccta agctacttaa acaagctgat
     2101 cctcaacaac aatctcctca ctggtcaaat cccaaaatct atcaagaatc tacaaaagct
     2161 tacacttctt gatcttagct acaacagctt atccggtgaa atcccacaag aactcggtca
     2221 agtcacaagc ttaactataa acctcgactt aagctacaac accttcactg gaaacatccc
     2281 ggaaaccttc tccgacctca cacagctaca atcactcgac ctctcaagca actcgctaca
     2341 cggcgacatc aaagtcctcg gttcactcac aagcctagct tcactaaaca tctcctgcaa
     2401 caacttctca ggtccaatcc catcaacacc attcttcaaa acaatctcca caacatcata
     2461 cctccaaaac acaaacctct gccattcact agacggcatc acttgctcat cacacaccgg
     2521 acaaaacaac ggagtaaaat ccctggaggt gctgttccag ggaccaagcg gcaatcccgg
     2581 cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg ccctgcagga
     2641 cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat
     2701 ggagtgcgac gcctgcggca tgcagccctc tagaggctcc ggctccatca tcccagttga
     2761 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     2821 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     2881 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     2941 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     3001 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     3061 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     3121 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     3181 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     3241 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     3301 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     3361 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     3421 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     3481 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     3541 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     3601 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     3661 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     3721 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     3781 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     3841 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     3901 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     3961 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     4021 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     4081 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     4141 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     4201 cgcgcacccg ggttctagag gaagcacctc cggcgataac gattacaagg atgatgatga
     4261 taaggccagc ggcaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     4321 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     4381 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4501 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4561 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     4621 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4681 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4741 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4801 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4861 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4921 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4981 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5041 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5101 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5161 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5221 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5281 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5341 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5401 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5461 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5521 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5581 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5641 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5701 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5761 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5821 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5881 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5941 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6001 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6061 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6121 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6181 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6241 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6301 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6361 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6421 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6481 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6541 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6601 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6661 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6721 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6781 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6841 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6901 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6961 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7021 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     7081 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     7141 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     7201 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
