LOCUS       sequence-298806-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector RGI5 X047_pECIA2 should be used with prey vector RGI5 
            X047_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6290)
  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 6290)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..6290
                     /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             3272..3313
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3323..3340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3363..3380)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3413..3547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3437..3456)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3491..3510
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3691..3707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3691..3707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3704..3726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3715..3731
                     /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(3739..3745,3746..3763,3764..3769))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3784..3805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3922..3939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4093..4681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4173..4192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4852..5643,5644..5712))
                     /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     5475..5494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5713..5817)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5885..5903
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5941..5963)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6063..6082
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6276..6290,1..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 ggaccaggga gcgacaaaac
     2581 tcacacatgc ccaccgtgcc cagcacctga actcctgggg ggaccgtcag tcttcctctt
     2641 ccccccaaaa cccaaggaca ccctcatgat ctcccggacc cctgaggtca catgcgtggt
     2701 ggtggacgtg agccacgaag accctgaggt caagttcaac tggtacgtgg acggcgtgga
     2761 ggtgcataat gccaagacaa agccgcggga ggagcagtac aacagcacgt accgtgtggt
     2821 cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca agtgcaaggt
     2881 ctccaacaaa gccctcccag cccccatcga gaaaaccatc tccaaagcca aagggcagcc
     2941 ccgagaacca caggtgtaca ccctgccccc atcccgggag gagatgacca agaaccaggt
     3001 cagcctgacc tgcctggtca aaggcttcta tcccagcgac atcgccgtgg agtgggagag
     3061 caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact ccgacggctc
     3121 cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg ggaacgtctt
     3181 ctcatgctcc gtgatgcatg agggtctgca caaccactac acgcagaaga gcctctccct
     3241 gtctccgggt aaatctagag ggcccttcga aggtaagcct atccctaacc ctctcctcgg
     3301 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     3361 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     3421 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3481 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3541 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3601 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3661 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5821 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5881 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5941 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6001 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     6061 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6121 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     6181 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6241 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
