LOCUS       sequence-270423-        6155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT4G34220 X018_pECIA14 should be used with bait vector 
            AT4G34220 X018_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6155)
  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 6155)
  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: AT4G34220 X018_pECIA14
FEATURES             Location/Qualifiers
     source          1..6155
                     /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             1451..1474
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3149..3172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3188..3205
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3228..3245)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3278..3412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3302..3321)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3356..3375
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3556..3572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3556..3572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3569..3591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3580..3596
                     /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(3604..3610,3611..3628,3629..3634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3649..3670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3787..3804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3958..4546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4038..4057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4717..5508,5509..5577))
                     /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     5340..5359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5578..5682)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5750..5768
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5806..5828)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5928..5947
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6141..6155,1..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 aacactgatg gtgttcttct
      541 gcttactttc aaatactcca tcctcactga ccctctctct gttctacgca attggaacta
      601 tgatgatgca acaccatgct tgtggacagg tgtcacatgt acagagcttg ggaagcctaa
      661 cacaccagac atgttcagag tcacaagctt ggtacttccc aacaaacacc ttttgggttc
      721 catcactccg gacttgttct cgattccata tctaaggatc ttggatctgt ctagcaattt
      781 cttcaacggg tcactcccgg actcggtgtt caacgccact gagcttcaga gtatctccct
      841 tggaagcaac aatctctctg gtgatttgcc caagagtgtc aacagtgtga ctaatctcca
      901 gctcttgaat ctctcagcta acgcatttac tggcgaaatt ccgctcaaca tctcacttct
      961 gaagaatcta acggtcgttt ccttgtcaaa aaacaccttt tcaggtgata ttccaagtgg
     1021 tttcgaagca gcacagattc ttgatctttc ttcaaatctt ctcaacggct ctcttcccaa
     1081 ggatttggga ggaaaaagcc tgcattactt gaacttatca cacaacaaag tcctcggcga
     1141 gatttctcca aattttgcag agaagtttcc agcaaatgcc acagtcgatc tctcctttaa
     1201 caacctcaca ggcccaatcc caagttccct ttctttactt aaccaaaagg ctgagtcttt
     1261 ttccggtaac caagagcttt gtgggaagcc attgaagatc ctttgttcaa ttccttccac
     1321 tctttcaaac cctccaaata tctctgaaac tacttcacca gcaatagcag ttaaaccaag
     1381 aagtacagct ccaataaacc ctttgacaga aaaaccaaat cagacaggaa aaagcaagct
     1441 aaagccaagc ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
     1501 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
     1561 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
     1621 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
     1681 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     1741 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     1801 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     1861 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     1921 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     1981 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     2041 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     2101 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     2161 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     2221 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
     2281 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     2341 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
     2401 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     2461 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     2521 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     2581 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     2641 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     2701 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     2761 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     2821 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     2881 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     2941 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     3001 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     3061 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     3121 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
     3181 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     3241 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     3301 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3361 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3421 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3481 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     3541 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3601 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3661 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3721 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3781 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3841 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3901 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3961 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4021 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4081 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4141 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4201 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4261 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4321 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4381 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4441 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4501 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4561 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4621 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4681 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4741 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4801 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4861 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4921 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4981 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5041 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5101 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5161 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5221 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5281 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5341 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5401 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5461 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5521 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5581 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5641 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5701 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5761 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5821 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5881 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5941 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6001 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6061 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6121 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
