LOCUS       sequence-227495-        5705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G05700 L9_pECIA2 should be used with prey vector 
            AT1G05700 L9_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5705)
  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 5705)
  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: AT1G05700 L9_pECIA2
FEATURES             Location/Qualifiers
     source          1..5705
                     /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             1958..1981
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2687..2728
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2738..2755
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2778..2795)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2828..2962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2852..2871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2906..2925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3106..3122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3106..3122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3119..3141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3130..3146
                     /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(3154..3160,3161..3178,3179..3184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3199..3220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3337..3354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3508..4096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3588..3607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4267..5058,5059..5127))
                     /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     4890..4909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5128..5232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5300..5318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5356..5378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5478..5497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5691..5705,1..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 tcaggtttca taagcatcga
      541 ttgtggaatt ccaagtggat catcgtacaa agacgacaca acaggtataa actacgtctc
      601 agattcatcc tttgtcgaaa ccggagtctc caaatcgatc cctttcacag ctcaaagaca
      661 gcttcaaaac ctgaggagct ttcccgaagg ttcaagaaac tgctatacat tgattccaat
      721 acaagggaag ggcaagaaat atctcataag agctagtttc atgtacggta actacgatgg
      781 agaaaatggt tcaccggagt ttgatctgtt tcttggtggc aatatttggg ataccgtctt
      841 gctcagcaat gggtcgagca ttgtttccaa agaagtcgtt tacttgagcc aatctgagaa
      901 catatttgtg tgtttgggaa acaaaggcaa aggaactcca tttatctcga ctttagagct
      961 taggtttctc gggaacgaca atacgacata tgattctcca aacggtgctc tcttcttctc
     1021 tagacgctgg gacttacgct ctctcatggg ttcacctgtt agatatgatg atgatgtgta
     1081 cgacagaatc tggatacctc gcaactttgg ctattgtaga gagatcaata cctcactccc
     1141 cgtgacctca gacaataata gttatagtct ttcaagtttg gtgatgagca cagcgatgac
     1201 tccaataaac acaacaagac ccatcacaat gactttggaa aacagtgatc caaatgtcag
     1261 gtactttgtc tacatgcact tcgcagaggt tgaagatctt agtctcaaac cgaaccagac
     1321 aagagagttc gacatcagca ttaacggagt gacagttgct gctggattca gccccaagta
     1381 tcttcagaca aacacgtttt ttctaaaccc tgagagccaa tcgaaaattg cgttttcact
     1441 cgttcgaacc ccaaagtcca ctcttccgcc aattgttaac gctctagaga tctacgtcgc
     1501 aaatagtttc tcgcagtctc tcactaacca agaggatggt gacgcggtta cgagtctaaa
     1561 gacgagttat aaagtgaaga agaactggca cggagatcct tgtttgccta atgactacat
     1621 ttgggagggc cttaattgca gttatgatag tcttactcct ccaagaatca catcactgaa
     1681 cttatcatca agcgggttaa cgggtcatat atcttcctcc ttctctaacc tcacaatgat
     1741 tcaagagcta gacttatcaa acaatggctt aaccggagac attccggagt tcctctcaaa
     1801 actgaagttc ttgagggttt taaatctaga aaataacact cttaccggtt cagttccatc
     1861 cgagttatta gaaagatcaa acaccggatc attctcgcta aggctaggag aaaatccagg
     1921 actctgtact gagatttctt gcagaaaaag caacagcctg gaggtgctgt tccagggacc
     1981 agggagcgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
     2041 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
     2101 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
     2161 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
     2221 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
     2281 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
     2341 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
     2401 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     2461 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     2521 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     2581 gcaggggaac gtcttctcat gctccgtgat gcatgagggt ctgcacaacc actacacgca
     2641 gaagagcctc tccctgtctc cgggtaaatc tagagggccc ttcgaaggta agcctatccc
     2701 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     2761 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     2821 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2881 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2941 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3001 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     3061 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3121 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3181 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3241 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5281 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5341 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5401 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5461 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5581 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5641 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     5701 acgtt
//
