LOCUS       sequence-274705-        6935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G34420 X003_pECIA14 should be used with bait vector 
            AT1G34420 X003_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6935)
  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 6935)
  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: AT1G34420 X003_pECIA14
FEATURES             Location/Qualifiers
     source          1..6935
                     /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             2231..2254
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3929..3952
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3968..3985
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4008..4025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4058..4192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4082..4101)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4136..4155
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4336..4352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4336..4352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4349..4371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4360..4376
                     /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(4384..4390,4391..4408,4409..4414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4429..4450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4567..4584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4738..5326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4818..4837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5497..6288,6289..6357))
                     /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     6120..6139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6358..6462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6530..6548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6586..6608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6708..6727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6921..6935,1..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 tcgttgaacc agacaaacac
      541 aatgatcgaa ctctctagtt tccttaatat ctcggattgg aatcttcctg gttctgagag
      601 gaatccatgt tcatggaacg gtgttctctg cagcttacca gacaatagct ctgtgattag
      661 cctgtctctc tctaactttg atctctccaa ttcttccttc ttaccacttg tctgcaatct
      721 ccagactttg gagtctcttg atgtctccaa caacagattg agctcaatcc cagaagggtt
      781 cgttacaaat tgcgaaagac tcattgctct gaagcatctg aacttcagca cgaacaagtt
      841 ttctacttcc cctggtttcc gcggtttctc caaattggcg gttcttgatt tctctcacaa
      901 cgtgttgagt gggaacgttg gagactatgg ttttgatggg ttggttcagt tgaggagttt
      961 gaatcttagc ttcaacagat taactggttc tgttccggtt catttgacca aaagtctaga
     1021 gaagcttgag gtttcagata atagtttgag cggtactatt cccgaaggca tcaaagatta
     1081 tcaagaacta acgctgatag atctcagtga taatcagctt aacggctcta tcccgagttc
     1141 gttgggaaac ctctccaagt tagaaagttt acttctctcc aacaactatc ttagtggact
     1201 aatcccagaa tcactttcaa gcatccagac attgcgtcgg tttgcagcga accgtaaccg
     1261 attcacggga gaaattccat cagggctcac aaaacacctc gagaacttag atctcagttt
     1321 caacagttta gcggggtcga ttccaggtga ccttttgtcg cagcttaaac tcgtctctgt
     1381 ggatctgtca tctaatcaac ttgttggatg gataccacaa agcatttcgt ctagcctggt
     1441 taggctaaga ctaggaagca ataagctaac agggtcagta ccctctgttg cgtttgaatc
     1501 gctgcagctc ctaacgtatc tggagatgga caacaacagt ttaactggct tcatacctcc
     1561 ttcatttgga aatcttgtga gcttgaatct cctgaacttg gctatgaacg agttcaccgg
     1621 gatcttacct cctgcatttg gaaaccttag caggcttcaa gtgataaaac tacagcaaaa
     1681 caagcttacg ggagaaatac cagatacaat cgcatttttg agtaacctct tgattcttaa
     1741 catcagctgc aattctctaa gtggatccat accgccttcg ctttctcagc taaagaggct
     1801 ttccaacatg aacttgcaag gcaacaatct gaatggtacc atacctgata acattcaaaa
     1861 tctggaggat ttgatagaac ttcagctggg acaaaaccag ctacgaggga ggatcccagt
     1921 aatgccacga aagttgcaaa tttccttaaa tctcagctac aacttgtttg aaggatctat
     1981 ccctactaca ttgtctgaac ttgaccgctt ggaagttctt gatctgtcaa ataacaactt
     2041 ctccggcgaa atacctaatt tcctgagcag attaatgtcc cttacacagc tcatactctc
     2101 caacaatcag cttaccggaa atattccaag gttcacccat aatgtttcag tcgatgtcag
     2161 aggaaaccct ggggttaagt tgaaaactga gaacgaagtc tctatccaga gaaatccatc
     2221 agggaaaagc ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
     2281 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
     2341 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
     2401 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
     2461 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     2521 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     2581 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     2641 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     2701 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     2761 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     2821 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     2881 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     2941 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     3001 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
     3061 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     3121 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
     3181 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     3241 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     3301 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     3361 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     3421 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     3481 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     3541 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     3601 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     3661 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     3721 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     3781 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     3841 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     3901 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
     3961 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     4021 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     4081 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4141 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4201 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4261 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     4321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4501 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5581 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5641 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5701 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5761 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5821 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5881 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5941 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6001 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6061 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6121 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6181 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6241 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6301 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6361 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6421 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6481 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6541 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6601 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6661 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6721 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6781 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6841 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6901 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
