LOCUS       sequence-297044-        5855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G37450 X025_pECIA2 should be used with prey vector 
            AT5G37450 X025_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5855)
  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 5855)
  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: AT5G37450 X025_pECIA2
FEATURES             Location/Qualifiers
     source          1..5855
                     /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             2108..2131
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2837..2878
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2888..2905
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2928..2945)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2978..3112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3002..3021)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3056..3075
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3256..3272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3256..3272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3269..3291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3280..3296
                     /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(3304..3310,3311..3328,3329..3334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3349..3370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3487..3504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3658..4246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3738..3757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4417..5208,5209..5277))
                     /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     5040..5059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5278..5382)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5450..5468
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5506..5528)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5628..5647
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5841..5855,1..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 actcatccca ctgatgttag
      541 tgctctgcag tatgttcacc gcaaactaaa ggaccctctg aatcatcttc aagactggaa
      601 aaagacggat ccatgcgcct ctaactggac cggtgtcatc tgcatccctg atccttctga
      661 tggctttctt catgttaaag aattgcggtt gctaaacatg aacctcactg gacaactggc
      721 acctgaactc ggcttactat caaatcttac catattgaac ttcatgtgga acgatctcac
      781 cggtcagatt ccaccagaac taggaaactt aactcatctc atcttcttgc tattaagtgg
      841 aaatcaactg acaggatctt tgcctcaaga actcggttct ctttccaatc tcttaatcct
      901 tcagatagat tacaatgaaa ttagcggaaa acttccaacg tccttggcaa atttgaaaaa
      961 actgaagcat tttcacatga acaataactc aatcacgggt cagattccgc ccgagtactc
     1021 taccttaacc aatgttctgc acttcttgat ggacaataac aagctaactg gtaatcttcc
     1081 accggagctc gctcagatgc caagcttgag aattctacaa ttggacggta gcaactttga
     1141 tggcacagag atcccatctt cttacggatc gataccaaat ctcgtcaaat tgagtctcag
     1201 aaactgcaac ctagaaggac caattcctga tttgagcaaa tcactggttc tctattactt
     1261 agatatttcc tccaacaaac tcacaggaga aattcccaag aacaagttct ctgcaaacat
     1321 cacaactata aacttgtaca acaatttgct cagcggatcc attccttcta atttttcagg
     1381 ccttccacgg ttacagagac tgcaagtgca gaacaacaac ttgagtgggg agattcctgt
     1441 gatatgggaa aataggatct tgaaggcaga ggaaaaactt atcttggact tgaggaacaa
     1501 catgttttca aatgtgtcaa gtgttcttct aaaccctccc tccaatgtca cagtcaagct
     1561 ttacggaaac ccggtttgtg caaatgttaa tgcgggaaag ttagctgatc tctgtggtat
     1621 ctcaacgtta gaagtggaat ctcccgcaac ttcttcggaa accatctcga cgggagactg
     1681 caaacgtcag tcgtgccccg taagcgaaaa ctatgactac gtgattgggt ctccagttgc
     1741 atgtttctgc gctgcaccac ttggcattga tcttcggctt aggagtccaa gtttctcgga
     1801 ctttcgacct tacaaagtct cctacatgct cgatgtagca tccccgaaaa atctcgggat
     1861 aaacccgtat cagatatcta ttgatacatt tgcatggcag tcgggtccaa ggctattcat
     1921 gaacatgaag attttccctg agtacagtga attaaacagt aaattcaaca gcacagaggt
     1981 tcaacgtatt gtggacttct ttgcaacttt cactctgaat actgatgatt ctcttggccc
     2041 ttatgagatt attagtatca acactggtgc ttacaaagac gttactatca tattcccgaa
     2101 aaagtcgctg gaggtgctgt tccagggacc agggagcgac aaaactcaca catgcccacc
     2161 gtgcccagca cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa
     2221 ggacaccctc atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca
     2281 cgaagaccct gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa
     2341 gacaaagccg cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt
     2401 cctgcaccag gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct
     2461 cccagccccc atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt
     2521 gtacaccctg cccccatccc gggaggagat gaccaagaac caggtcagcc tgacctgcct
     2581 ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga
     2641 gaacaactac aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag
     2701 caagctcacc gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat
     2761 gcatgagggt ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaatc
     2821 tagagggccc ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     2881 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     2941 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     3001 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3061 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3121 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3181 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     3241 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3301 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3361 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3421 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3481 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3541 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3601 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3661 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3721 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3781 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3841 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3901 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3961 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4021 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4081 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4141 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4201 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4261 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4321 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4381 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4441 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4501 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4561 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4621 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4681 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4741 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4801 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4861 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4921 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4981 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5041 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5101 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5161 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5221 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5281 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5341 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5401 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5461 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5521 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5581 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5641 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5701 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5761 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5821 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
