LOCUS       sequence-238693-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G06840 X062_pECIA14 should be used with bait vector 
            AT1G06840 X062_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  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 6812)
  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: AT1G06840 X062_pECIA14
FEATURES             Location/Qualifiers
     source          1..6812
                     /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             2553..2576
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4251..4274
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4290..4307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4330..4347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4380..4514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4404..4423)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4458..4477
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4658..4674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4658..4674)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4671..4693)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4682..4698
                     /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(4706..4712,4713..4730,4731..4736))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4751..4772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4889..4906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5060..5648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5140..5159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5819..6610,6611..6679))
                     /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     6442..6461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6680..6784)
                     /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 tcgggcaaga cgacatcacc aatcccgttg aagtgagggc tttgcgagtg atcaaagaaa
     1021 gtttgaacga tcctgttcat agattgagaa attggaagca tggagacccg tgcaattcga
     1081 attggactgg tgttgtctgc ttcaactcca ctcttgatga tggttatctt catgtcagcg
     1141 aattgcaatt gttcagtatg aatctctcag ggaacttgtc tccggagctt ggccggttgt
     1201 ctcgtcttac tatcctgagt ttcatgtgga ataagatcac cggaagtata ccaaaggaaa
     1261 ttgggaatat caagtcctta gaactcttgc tcctgaatgg aaatctgtta aatggaaact
     1321 taccagagga gctagggttt cttccaaact tggacagaat acagattgat gaaaaccgta
     1381 tatcaggacc acttcccaaa tcttttgcaa acttaaacaa aacgaagcac tttcacatga
     1441 acaataattc tattagcggg cagataccac cagagcttgg aagcctacca tccattgttc
     1501 acatccttct tgataacaac aatttatcag gctatcttcc ccctgagtta tcaaacatgc
     1561 cgcgtttact tatcctacaa ttagataaca atcactttga cgggactacg attccacaat
     1621 cttatggaaa catgtctaaa cttttgaaga tgagtcttag gaactgcagc ttgcaagggc
     1681 cagtgcctga tcttagcagc ataccgaacc ttggttattt ggacctaagt caaaatcagt
     1741 taaatggatc tatacctgca gggaagcttt ctgatagtat cacaaccatc gatctatcca
     1801 ataacagtct aactggaacc attccaacaa atttctcagg ccttccacgg cttcaaaagc
     1861 tgtcgcttgc aaacaatgct ctgagtggtt ccattccctc tagaatatgg caggaaagag
     1921 agctgaattc aaccgagagt attatcgtgg atctgcggaa caatgggttt tcaaatatct
     1981 ctggcagatc cgatctccgt ccaaatgtga ccgtctggct tcaggggaat ccgttgtgct
     2041 cagatggaaa tctgcttcga ttgtgtggac ctataactga ggaagacatt aatcagggtt
     2101 caaccaattc taatactaca atttgttctg actgcccacc cccttatgaa ttttcaccag
     2161 aacctcttag acgttgcttt tgtgctgctc ctctgcttgt tggatatcgg ttgaaaagtc
     2221 ctggtttctc ggactttgtt ccttacagat ccgaatttga gcaatatatc acctctggtc
     2281 ttagtttgaa tctgtatcag ctacgccttg actcattcca gtggcagaaa ggacctagac
     2341 ttcgaatgta tttgaagttc tttcctgttt ttggttcaaa tgccaacaat tctttcatat
     2401 tcaatcgtag cgaggttcgg cgaataaggg gcatgttcac tggatggaat atccgagacg
     2461 aagatctctt tggtccttat gagcttatga atttcacatt gttagatgtc tacagagatg
     2521 tgtttccttc agcttcgcca tctggtctaa gtctggaggt gctgttccag ggaccaagcg
     2581 gcaatcccgg cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg
     2641 ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga
     2701 acaccgtgat ggagtgcgac gcctgcggca tgcagccctc tagaggctcc ggctccatca
     2761 tcccagttga ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg
     2821 ccgccaagaa gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg
     2881 atgggatggg ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca
     2941 aactggggcc tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga
     3001 catacaatgt agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg
     3061 gggtcaaggg caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca
     3121 acacgacacg cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt
     3181 cagtgggagt ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc
     3241 acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg
     3301 ggtgccagga catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg
     3361 gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca
     3421 gccaaggtgg gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc
     3481 agggtgcccg gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg
     3541 tgacccatct catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact
     3601 ccacactgga cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga
     3661 acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa
     3721 gcagggctta ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg
     3781 gccagctcac cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct
     3841 tctccttcgg aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg
     3901 cccgggacag gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca
     3961 aggacggcgc ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc
     4021 agtcagcagt gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc
     4081 gcggcccgca ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca
     4141 tggccttcgc cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca
     4201 ccaccgacgc cgcgcacccg ggttctagag gaagcacctc cggcgataac gattacaagg
     4261 atgatgatga taaggccagc ggcaccggtc atcatcacca tcaccattga gtttaaaccc
     4321 gctgatcagc ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg
     4381 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     4441 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     4501 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     4561 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
     4621 agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4681 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4741 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4801 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4861 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4921 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4981 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5461 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5581 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5641 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5701 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5761 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5821 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5881 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5941 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6001 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6061 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6121 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6181 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6241 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6301 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6361 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6421 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6481 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6541 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6601 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6661 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6721 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6781 cgcgcacatt tccccgaaaa gtgccacctg ac
//
