LOCUS       sequence-274536-        6962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G53440 X069_pECIA14 should be used with bait vector 
            AT1G53440 X069_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6962)
  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 6962)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..6962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     330..383
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2121..2144
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3819..3842
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3858..3875
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3898..3915)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3948..4082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3972..3991)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4026..4045
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4226..4242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4226..4242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4239..4261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4250..4266
                     /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(4274..4280,4281..4298,4299..4304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4319..4340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4457..4474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4628..5216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4708..4727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5387..6178,6179..6247))
                     /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     6010..6029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6248..6352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6420..6438
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6476..6498)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6598..6617
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6811..6833
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6826..6842
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(6848..6962,1..312)
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
ORIGIN
        1 acccccatac atatgtggta cgcaagtaag agtgcctgcg catgccccat gtgccccacc
       61 aagagctttg catcccatac aagtccccaa agtggagaac cgaaccaatt cttcgcgggc
      121 agaacaaaag cttctgcaca cgtctccact cgaatttgga gccggccggc gtgtgcaaaa
      181 gaggtgaatc gaacgaaaga cccgtgtgta aagccgcgtt tccaaaatgt ataaaaccga
      241 gagcatctgg ccaatgtgca tcagttgtgg tcagcagcaa aatcaagtga atcatctcag
      301 tgcaactaaa ggggggatcc gatctcaata tgaagttatg catattactg gccgtcgtgg
      361 cctttgttgg cctctcgctc gggcaacttt tgccagaaga tgaagttcaa acattgcgaa
      421 cgatctttag aaagcttcaa aaccaaacag tgaacatcga aagaacttct tgtttggaca
      481 gaaaatggaa ctttgttgcc gagtcaacct ccaagctacc aaccagtaac attacctgcg
      541 actgtacttt caacgccagc tcagtctgtc gtgtcacaaa catacagctt agaggtttca
      601 atttgcgagg aattataccg cctgaattcg ggaacctcac acgtcttaca gagatagatc
      661 ttgtgctgaa ctttctcagt ggaacaatac ctacaacatt gtctcaaatt ccgcttgaaa
      721 tcttggctgt aaccggaaac cgactctctg gaccatttcc tcctcagctc ggacagatta
      781 ctacacttac tgatgtgatt atggaaagta atttattcac aggacaactt cctccaaacc
      841 tagggaactt aagaagcttg aaaagattgc taatctcttc aaataacatc acgggtcgga
      901 tccctgagtc cttgagcaat ctcaagaatt tgactaattt tcggattgat ggaaactctc
      961 tatctgggaa gatacctgat tttattggaa actggactcg gctcgttagg ctagacctcc
     1021 aaggcacatc aatggaaggt cctattccag cttcgatttc aaacttgaaa aacctgactg
     1081 aattgaggat aacagatttg cgtggaccaa cttctccttt tccagacctt caaaatatga
     1141 cgaatatgga aagattggta ctaagaaact gtttgataag ggaacctata ccggagtaca
     1201 tcggtacttc catgactatg ttgaagttac tagatttaag ctcaaacatg ttaaatggta
     1261 caattccaga cacatttcgg agtctgaacg cattcaactt tatgtatctg aataataact
     1321 cattgactgg tccagttcct cagttcattc ttgatagtaa acaaaacata gatttatctt
     1381 acaacaattt cactcagcca cctactttaa gctgtaatca gcttgatgtt aacttgatct
     1441 ccagctaccc ctcagtaacc aataactctg tccaatggtg cttaagaaag gatcttccgt
     1501 gtcctggaga cgcacaccat tcttccttgt tcattaactg tggaggaaac cgactcaagg
     1561 ttgataaaga cgagtatgcg gatgacttaa acaaaagagg agcatcaaca ttctcttctg
     1621 tctctgaaag atggggatac agtagttctg gagcttggtt aggcaatgac ggtgccactt
     1681 acttagcaac agatacgttt aacttgatca atgaatcaac tccagagtat tacaaaacag
     1741 cccgtctcgc ttcacaatca ctcaagtact atggactatg catgagaaga ggaagttaca
     1801 aagttcagct ctattttgca gagataatgt tctcaaatga tcagacttat agtagcttag
     1861 ggcggcgact attcgacatt tatgttcaag ggattttgtt ggagagggac tttaacatag
     1921 cacagagagc aggtggagtt ggtaaaccgt tcctaaggca agttgatgaa gttcaagtga
     1981 atggaagtac gttggagatt catttgaagt ggacaggaaa aggcacaaac gtaataccaa
     2041 caagaggtgt ttacgggcct ctcatatccg ctataaccgt cacaccgaat ttcaaggttg
     2101 ataccggaaa accattgtcc ctggaggtgc tgttccaggg accaagcggc aatcccggcg
     2161 gcgatctggc cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg
     2221 tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg
     2281 agtgcgacgc ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg
     2341 aggagaaccc ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc
     2401 tgcagcctgc acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg
     2461 tgtctacggt gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg
     2521 agatacccct ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag
     2581 acaaacatgt gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca
     2641 acttccagac cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg
     2701 gcaacgaggt catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg
     2761 taaccaccac acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc
     2821 gcaactggta ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca
     2881 tcgctacgca gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt
     2941 acatgtttcg catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga
     3001 ccaggctgga cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt
     3061 atgtgtggaa ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca
     3121 tgggtctctt tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc
     3181 cctccctgat ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct
     3241 tcttcctctt cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc
     3301 gggcactgac tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca
     3361 gcgaggagga cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag
     3421 gctaccccct gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga
     3481 aggcctacac ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc
     3541 ggccggatgt taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc
     3601 ccctggacga agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg
     3661 cgcacctggt tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg
     3721 cctgcctgga gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg
     3781 cgcacccggg ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata
     3841 aggccagcgg caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     3901 cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga
     3961 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4021 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4081 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4141 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca
     4201 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4261 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4321 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4381 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4801 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5041 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6361 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6421 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6481 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6541 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6601 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6661 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     6721 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6781 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6841 gtgaattaat tcgttgcagg acaggatgtg gtgcccgatg tgactagctc tttgctgcag
     6901 gccgtcctat cctctggttc cgataagaga cccagaactc cggcccccca ccgcccaccg
     6961 cc
//
