LOCUS       sequence-238698-        6980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G56120 X070_pECIA14 should be used with bait vector 
            AT1G56120 X070_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6980)
  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 6980)
  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: AT1G56120 X070_pECIA14
FEATURES             Location/Qualifiers
     source          1..6980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2721..2744
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4419..4442
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4458..4475
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4498..4515)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4548..4682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4572..4591)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4626..4645
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4826..4842)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4826..4842)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4839..4861)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4850..4866
                     /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(4874..4880,4881..4898,4899..4904))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4919..4940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5057..5074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5228..5816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5308..5327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5987..6778,6779..6847))
                     /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     6610..6629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6848..6952)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggtccat cttcgcggct tggaagattc gggcgccgag ggaatggaac atcagcggcg
     1021 aactttgctc cggcgtcgcc atcgacgcca gtgtactcga ctcaaaccat gcctacaatc
     1081 ctctcatcaa atgcgactgt agtttccaaa actccacaat ctgccgcata aataacataa
     1141 aggtttatgc gatagatgtt gtaggaccta tacctccaga gctctggacc ttaacatacc
     1201 tcaccaatct gaacctaggt caaaattatc tcacaggctc actgtctcct gctattggga
     1261 atttgactcg gatgcagtgg atgacttttg gaatcaacgc gttgtctggc cctattccta
     1321 aggaaatcgg tttgcttacg gatttaagat tacttggtat tagttcaaat aacttttccg
     1381 gttccttacc agctgagatt gggagttgta caaaactaca gcaaatgtac atcgatagtt
     1441 ctggactcag cggaggaata cctttatcat ttgctaattt tgtggagttg gaagtcgctt
     1501 ggattatgga tgtggaactt acaggtcgga taccagactt tataggattc tggaccaaac
     1561 ttactacctt gaggatcctc ggaactggtt tgagcggccc aataccgtcg tcattttcca
     1621 acttaattgc tttgacagaa ctgaggcttg gtgatatatc caatggaagc tcttctcttg
     1681 atttcatcaa agacatgaaa tctctaagtg tattagtatt gaggaacaac aatctcactg
     1741 ggacaatacc gtctactatt ggaggataca caagtttgca acaagttgat ttgagcttca
     1801 acaaattaca tggaccaatt ccggcttcac ttttcaactt aagtcgactt actcacttgt
     1861 ttctgggaaa caacacgttg aatggatcct tgcccactct aaaggggcag tctctgagca
     1921 atttagatgt gtcgtacaat gatttgtctg gaagtcttcc ttcatgggtc agcttaccag
     1981 atttgaagct caatctagtt gctaacaact tcacactgga aggtcttgac aacagggttt
     2041 tatcaggact acattgcctg cagaagaact tcccctgcaa tcgaggcgaa ggaatctatt
     2101 ataacttttc gatcaactgc ggaggcccag agataaggtc tgttagtgga gcactatttg
     2161 aaaaggagga cgcggatctt ggaccagctt cgtttgttgt gagtgctgca aagagatggg
     2221 cagccagtag tgtaggaaat tttgctggca gtagcaacaa tatatacata gctacttcac
     2281 tagcacaatt tatcaacact atggactcag agctttttca gtcagcacga ctttctgcat
     2341 cttccctaag gtattatggg ttggggctag aaaatggagg gtacaccgta acacttcagt
     2401 ttgctgaagt acaaattgaa ggttctaact cttggaaagg tattggaaga cgacgtttta
     2461 acatttatgt ccagggaaga cttgttgaaa aggattttga tatacgcaga acagctggtg
     2521 gctcttcggt tcgagcagtt cagagagaat ataaaacaaa tgtatcagaa aatcatctcg
     2581 aagtccatct tttctgggct ggaaaaggaa catgttgtat tcctatccaa ggggcttatg
     2641 ggccattaat agcggccgtc agtgcgacac cagatttcac accaactgtg gctaataggc
     2701 caccatcaaa gggaaagagc ctggaggtgc tgttccaggg accaagcggc aatcccggcg
     2761 gcgatctggc cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg
     2821 tgcgcgagct gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg
     2881 agtgcgacgc ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg
     2941 aggagaaccc ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc
     3001 tgcagcctgc acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg
     3061 tgtctacggt gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg
     3121 agatacccct ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag
     3181 acaaacatgt gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca
     3241 acttccagac cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg
     3301 gcaacgaggt catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg
     3361 taaccaccac acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc
     3421 gcaactggta ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca
     3481 tcgctacgca gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt
     3541 acatgtttcg catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga
     3601 ccaggctgga cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt
     3661 atgtgtggaa ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca
     3721 tgggtctctt tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc
     3781 cctccctgat ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct
     3841 tcttcctctt cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc
     3901 gggcactgac tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca
     3961 gcgaggagga cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag
     4021 gctaccccct gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga
     4081 aggcctacac ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc
     4141 ggccggatgt taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc
     4201 ccctggacga agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg
     4261 cgcacctggt tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg
     4321 cctgcctgga gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg
     4381 cgcacccggg ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata
     4441 aggccagcgg caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     4501 cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga
     4561 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4621 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4681 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4741 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca
     4801 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4861 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4921 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4981 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5041 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5101 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5161 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5221 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5281 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5341 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5401 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5461 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5521 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5581 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5641 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5701 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5761 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5821 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5881 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5941 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6001 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6061 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6121 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6181 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6241 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6301 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6361 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6421 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6481 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6541 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6601 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6661 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6721 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6781 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6841 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6901 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6961 cccgaaaagt gccacctgac
//
