LOCUS       sequence-296638-        5708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G46420 N10_pECIA2 should be used with prey vector 
            AT3G46420 N10_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  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 5708)
  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: AT3G46420 N10_pECIA2
FEATURES             Location/Qualifiers
     source          1..5708
                     /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             1961..1984
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2690..2731
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2741..2758
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2781..2798)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2831..2965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2855..2874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2909..2928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3109..3125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3109..3125)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3122..3144)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3133..3149
                     /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(3157..3163,3164..3181,3182..3187))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3202..3223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3340..3357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3511..4099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3591..3610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4270..5061,5062..5130))
                     /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     4893..4912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5131..5235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5303..5321
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5359..5381)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5481..5500
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5694..5708,1..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 ggatttatca gtgtagactg
      541 cggattatct cctaatgagg tctctcctta tattgagccg tttactggac tacagttcac
      601 aacggattca aatttcattg aaaccggaaa acttggtaga attcaggcaa gcctagagcc
      661 gaagtaccga aagtcgcaaa cgacgttgag atattttcct gatggaatac gcaactgtta
      721 caatcttaca gtgacacaag gaaccaacta tttaatcaga gctagagcaa tatatgggaa
      781 ttatgatggt cttaacattt atcctaaatt tgacttgtac ataggcccta atttttgggt
      841 aacaatagac ttaggaaagt atgtaaatgg tacatgggag gagatcatct acataccaaa
      901 atctaatatg ttggacgtgt gtcttgtcaa gacagggccg tccacaccat tgatatcatc
      961 cttggtgtta aggcctcttg caaatgctac ttacatcact caatctggtt ggttgaagac
     1021 ttatgttcgg gtttatctaa gcgactcaaa tgacgtcata cgatatccgg atgatgtcta
     1081 tgatcgtata tggggttcat actttgagcc ggagtggaag aagatttcca ctacacttgg
     1141 agtgaacagt tctagtggat ttctgcctcc actaaaggca ctcatgactg ccgcctctcc
     1201 tgccaatgct agtgcaccat tggctattcc cggagtacta gatttcccaa gtgataaact
     1261 atacttgttc ctccacttct ctgagattca agtcttaaag gccaacgaaa ctagagagtt
     1321 cgaaattttt tggaataaaa agcttgttta caatgcttat agtcctgtat atctacagac
     1381 caagactata cgcaacccat ctccagttac ttgtgaaaga ggagaatgca tactagagat
     1441 gatcaaaact gagaggtcaa ctctcccgcc tttgcttaac gccgttgaag ttttcacagt
     1501 ggttgaattt cctcagccgg aaacagatgc tagtgatgtg gtggctataa aaaacatcaa
     1561 ggccatctat ggattgactc gagtcacatg gcaaggagat ccatgtgttc ctcaacaatt
     1621 tttgtggaac ggtttaaatt gcaacagcat ggagacgtct acacctccaa gaatcacttc
     1681 cttggacttg tcttcaagcg ggttaaccgg aagcatatca gttgttattc aaaatcttac
     1741 ccatctagaa aagttggatt tgtcaaataa caacttgact ggggaagttc ccgattttct
     1801 agcaaacatg aaatttcttg tgttcataaa cttaagcaag aacaatctga atggttcgat
     1861 tcctaaagct cttcgcgata gagaaaataa aggacttaag ttaattgtcg acaaaaatgt
     1921 ggacaactgc tcatctggtt catgcaccca aaaaaagaaa ctggaggtgc tgttccaggg
     1981 accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     2041 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     2101 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     2161 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     2221 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     2281 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     2341 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     2401 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     2461 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     2521 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     2581 gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca accactacac
     2641 gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag gtaagcctat
     2701 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     2761 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     2821 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2881 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2941 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3001 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3061 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3121 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3181 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3241 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3301 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3361 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3421 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5281 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5341 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5401 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5461 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5521 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5581 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5641 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5701 acgacgtt
//
