LOCUS       sequence-274067-        7055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G28040 X014_pECIA14 should be used with bait vector 
            AT3G28040 X014_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7055)
  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 7055)
  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: AT3G28040 X014_pECIA14
FEATURES             Location/Qualifiers
     source          1..7055
                     /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             2351..2374
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4049..4072
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4088..4105
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4128..4145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4178..4312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4202..4221)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4256..4275
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4456..4472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4456..4472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4469..4491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4480..4496
                     /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(4504..4510,4511..4528,4529..4534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4549..4570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4687..4704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4858..5446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4938..4957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5617..6408,6409..6477))
                     /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     6240..6259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6478..6582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6650..6668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6706..6728)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6828..6847
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7041..7055,1..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 tcaattcaac taaacgacga
      541 cgttttagga ctcatcgtct tcaaatcaga cctcaacgac ccattttcac accttgagtc
      601 atggaccgaa gacgacaaca ctccttgttc atggagttac gtcaaatgca accccaaaac
      661 atctcgagtc atcgagcttt ctctcgacgg tttagcctta accggaaaaa tcaaccgcgg
      721 aatccaaaag cttcaacgtt taaaagtact ctcactttcc aacaacaact tcaccggaaa
      781 catcaacgct ctctcaaaca acaaccatct ccaaaagcta gatcttagcc acaacaatct
      841 ctccggtcaa atcccttctt ctctcggttc aataacctcc ttgcaacacc ttgacttaac
      901 cggaaactcc ttctccggta cactctctga cgatttgttc aacaactgtt cctctcttag
      961 atatctttct ctctcccaca accacctcga aggtcaaatc ccaagtactc tgtttcgatg
     1021 ctctgtttta aacagtctca atctttcaag aaaccgtttc tccggtaacc ctagcttcgt
     1081 ctcaggaatc tggagacttg agaggttaag agctttagat ctatcttcta actcactttc
     1141 tggttcaata cctttgggga tactctctct acataacttg aaagagttgc aactacagag
     1201 gaatcagttc tctggagcat tgccttcaga tattggactc tgtcctcatt taaacagagt
     1261 tgatctaagt tccaatcatt tctctggtga acttccaaga actcttcaga agctgaaatc
     1321 tttaaatcac ttcgatgtat cgaacaattt gctctccggt gatttcccgc catggatcgg
     1381 tgacatgacc ggtttagtac acttggattt ctccagcaat gagttaaccg gaaagcttcc
     1441 ttcttctatt agtaacttga ggtctctaaa ggatctaaac ttgtctgaga acaaactctc
     1501 cggcgaggta cctgagtctt tggaatcatg caaagagctt atgattgttc agcttaaagg
     1561 caatgacttc tccggtaaca ttcctgatgg tttttttgat cttggtcttc aagaaatgga
     1621 tttttcgggt aacggtttaa ccggttcgat cccaagaggc tcaagcaggc ttttcgagtc
     1681 actcataagg ctcgatcttt cgcataacag tctcactgga agtatacctg gtgaagtagg
     1741 actcttcatc cacatgagat accttaattt atcatggaac catttcaaca caagagttcc
     1801 tcctgaaatt gagtttctac agaacttaac ggtattggat cttaggaaca gtgcactgat
     1861 tggttcggtt cctgctgata tatgtgagtc tcagagtctt cagatacttc aactggatgg
     1921 taactcacta accggttcta taccggaagg aatcggaaac tgctcttctc ttaaattgtt
     1981 gagtttgtct cataacaatc ttaccggtcc tattcctaaa tctctttcaa acttacaaga
     2041 gctcaagatt ttaaagctag aggctaataa gcttagtgga gaaataccga aagagcttgg
     2101 ggatttgcag aatctgttat tggttaacgt ttcgtttaac cgacttatcg gaaggttacc
     2161 attgggagat gtgttccaaa gcttagacca gagtgctata caaggaaacc taggtatttg
     2221 ttcaccgttg ttgagaggtc cttgtacact gaatgttcca aagcctcttg tcatcaatcc
     2281 aaactcctac gggaatggga acaatatgcc tggaaaccga gcaagcggtg gttctggaac
     2341 attccaccgc ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
     2401 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
     2461 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
     2521 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
     2581 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     2641 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     2701 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     2761 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     2821 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     2881 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     2941 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     3001 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     3061 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     3121 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
     3181 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     3241 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
     3301 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     3361 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     3421 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     3481 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     3541 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     3601 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     3661 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     3721 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     3781 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     3841 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     3901 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     3961 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     4021 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
     4081 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     4141 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     4201 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4321 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4381 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     4441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5041 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5281 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6601 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6661 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6721 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6781 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6841 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6901 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6961 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7021 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
