LOCUS       sequence-297038-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G53440 X069_pECIA2 should be used with prey vector 
            AT1G53440 X069_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  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 6005)
  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: AT1G53440 X069_pECIA2
FEATURES             Location/Qualifiers
     source          1..6005
                     /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             2258..2281
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2987..3028
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3038..3055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3078..3095)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3128..3262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3152..3171)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3206..3225
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3406..3422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3406..3422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3419..3441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3430..3446
                     /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(3454..3460,3461..3478,3479..3484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3499..3520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3637..3654)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3808..4396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3888..3907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4567..5358,5359..5427))
                     /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     5190..5209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5428..5532)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5600..5618
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5656..5678)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5778..5797
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5991..6005,1..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 caacttttgc cagaagatga
      541 agttcaaaca ttgcgaacga tctttagaaa gcttcaaaac caaacagtga acatcgaaag
      601 aacttcttgt ttggacagaa aatggaactt tgttgccgag tcaacctcca agctaccaac
      661 cagtaacatt acctgcgact gtactttcaa cgccagctca gtctgtcgtg tcacaaacat
      721 acagcttaga ggtttcaatt tgcgaggaat tataccgcct gaattcggga acctcacacg
      781 tcttacagag atagatcttg tgctgaactt tctcagtgga acaataccta caacattgtc
      841 tcaaattccg cttgaaatct tggctgtaac cggaaaccga ctctctggac catttcctcc
      901 tcagctcgga cagattacta cacttactga tgtgattatg gaaagtaatt tattcacagg
      961 acaacttcct ccaaacctag ggaacttaag aagcttgaaa agattgctaa tctcttcaaa
     1021 taacatcacg ggtcggatcc ctgagtcctt gagcaatctc aagaatttga ctaattttcg
     1081 gattgatgga aactctctat ctgggaagat acctgatttt attggaaact ggactcggct
     1141 cgttaggcta gacctccaag gcacatcaat ggaaggtcct attccagctt cgatttcaaa
     1201 cttgaaaaac ctgactgaat tgaggataac agatttgcgt ggaccaactt ctccttttcc
     1261 agaccttcaa aatatgacga atatggaaag attggtacta agaaactgtt tgataaggga
     1321 acctataccg gagtacatcg gtacttccat gactatgttg aagttactag atttaagctc
     1381 aaacatgtta aatggtacaa ttccagacac atttcggagt ctgaacgcat tcaactttat
     1441 gtatctgaat aataactcat tgactggtcc agttcctcag ttcattcttg atagtaaaca
     1501 aaacatagat ttatcttaca acaatttcac tcagccacct actttaagct gtaatcagct
     1561 tgatgttaac ttgatctcca gctacccctc agtaaccaat aactctgtcc aatggtgctt
     1621 aagaaaggat cttccgtgtc ctggagacgc acaccattct tccttgttca ttaactgtgg
     1681 aggaaaccga ctcaaggttg ataaagacga gtatgcggat gacttaaaca aaagaggagc
     1741 atcaacattc tcttctgtct ctgaaagatg gggatacagt agttctggag cttggttagg
     1801 caatgacggt gccacttact tagcaacaga tacgtttaac ttgatcaatg aatcaactcc
     1861 agagtattac aaaacagccc gtctcgcttc acaatcactc aagtactatg gactatgcat
     1921 gagaagagga agttacaaag ttcagctcta ttttgcagag ataatgttct caaatgatca
     1981 gacttatagt agcttagggc ggcgactatt cgacatttat gttcaaggga ttttgttgga
     2041 gagggacttt aacatagcac agagagcagg tggagttggt aaaccgttcc taaggcaagt
     2101 tgatgaagtt caagtgaatg gaagtacgtt ggagattcat ttgaagtgga caggaaaagg
     2161 cacaaacgta ataccaacaa gaggtgttta cgggcctctc atatccgcta taaccgtcac
     2221 accgaatttc aaggttgata ccggaaaacc attgtccctg gaggtgctgt tccagggacc
     2281 agggagcgac aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc
     2341 gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga
     2401 ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta
     2461 cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag
     2521 cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
     2581 gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa
     2641 agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc gggaggagat
     2701 gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc
     2761 cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct
     2821 ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
     2881 gcaggggaac gtcttctcat gctccgtgat gcatgagggt ctgcacaacc actacacgca
     2941 gaagagcctc tccctgtctc cgggtaaatc tagagggccc ttcgaaggta agcctatccc
     3001 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     3061 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     3121 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3181 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3241 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3301 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     3361 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     3421 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3481 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3541 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3601 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3661 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3721 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3781 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3841 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3901 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3961 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4021 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4081 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4141 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4201 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4261 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4321 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4381 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4441 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4501 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4561 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4621 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4681 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4741 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4801 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4861 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4921 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4981 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5041 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5101 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5161 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5221 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5281 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5341 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5401 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5461 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5521 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5581 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5641 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5701 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5761 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     5821 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     5881 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     5941 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6001 acgtt
//
