LOCUS       sequence-297014-        6098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G28040 X014_pECIA2 should be used with prey vector 
            AT3G28040 X014_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  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 6098)
  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: AT3G28040 X014_pECIA2
FEATURES             Location/Qualifiers
     source          1..6098
                     /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             2351..2374
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3080..3121
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3131..3148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3171..3188)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3221..3355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3245..3264)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3299..3318
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3499..3515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3499..3515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3512..3534)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3523..3539
                     /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(3547..3553,3554..3571,3572..3577))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3592..3613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3730..3747)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3901..4489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3981..4000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4660..5451,5452..5520))
                     /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     5283..5302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5521..5625)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5693..5711
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5749..5771)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5871..5890
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6084..6098,1..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 tcaattcaac taaacgacga
      541 cgttttagga ctcatcgtct tcaaatcaga cctcaacgac ccattttcac accttgagtc
      601 atggaccgaa gacgacaaca ctccttgttc atggagttac gtcaaatgca accccaaaac
      661 atctcgagtc atcgagcttt ctctcgacgg tttagcctta accggaaaaa tcaaccgcgg
      721 aatccaaaag cttcaacgtt taaaagtact ctcactttcc aacaacaact tcaccggaaa
      781 catcaacgct ctctcaaaca acaaccatct ccaaaagcta gatcttagcc acaacaatct
      841 ctccggtcaa atcccttctt ctctcggttc aataacctcc ttgcaacacc ttgacttaac
      901 cggaaactcc ttctccggta cactctctga cgatttgttc aacaactgtt cctctcttag
      961 atatctttct ctctcccaca accacctcga aggtcaaatc ccaagtactc tgtttcgatg
     1021 ctctgtttta aacagtctca atctttcaag aaaccgtttc tccggtaacc ctagcttcgt
     1081 ctcaggaatc tggagacttg agaggttaag agctttagat ctatcttcta actcactttc
     1141 tggttcaata cctttgggga tactctctct acataacttg aaagagttgc aactacagag
     1201 gaatcagttc tctggagcat tgccttcaga tattggactc tgtcctcatt taaacagagt
     1261 tgatctaagt tccaatcatt tctctggtga acttccaaga actcttcaga agctgaaatc
     1321 tttaaatcac ttcgatgtat cgaacaattt gctctccggt gatttcccgc catggatcgg
     1381 tgacatgacc ggtttagtac acttggattt ctccagcaat gagttaaccg gaaagcttcc
     1441 ttcttctatt agtaacttga ggtctctaaa ggatctaaac ttgtctgaga acaaactctc
     1501 cggcgaggta cctgagtctt tggaatcatg caaagagctt atgattgttc agcttaaagg
     1561 caatgacttc tccggtaaca ttcctgatgg tttttttgat cttggtcttc aagaaatgga
     1621 tttttcgggt aacggtttaa ccggttcgat cccaagaggc tcaagcaggc ttttcgagtc
     1681 actcataagg ctcgatcttt cgcataacag tctcactgga agtatacctg gtgaagtagg
     1741 actcttcatc cacatgagat accttaattt atcatggaac catttcaaca caagagttcc
     1801 tcctgaaatt gagtttctac agaacttaac ggtattggat cttaggaaca gtgcactgat
     1861 tggttcggtt cctgctgata tatgtgagtc tcagagtctt cagatacttc aactggatgg
     1921 taactcacta accggttcta taccggaagg aatcggaaac tgctcttctc ttaaattgtt
     1981 gagtttgtct cataacaatc ttaccggtcc tattcctaaa tctctttcaa acttacaaga
     2041 gctcaagatt ttaaagctag aggctaataa gcttagtgga gaaataccga aagagcttgg
     2101 ggatttgcag aatctgttat tggttaacgt ttcgtttaac cgacttatcg gaaggttacc
     2161 attgggagat gtgttccaaa gcttagacca gagtgctata caaggaaacc taggtatttg
     2221 ttcaccgttg ttgagaggtc cttgtacact gaatgttcca aagcctcttg tcatcaatcc
     2281 aaactcctac gggaatggga acaatatgcc tggaaaccga gcaagcggtg gttctggaac
     2341 attccaccgc ctggaggtgc tgttccaggg accagggagc gacaaaactc acacatgccc
     2401 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     2461 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     2521 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     2581 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     2641 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     2701 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     2761 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     2821 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     2881 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     2941 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     3001 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     3061 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     3121 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     3181 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     3241 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3301 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3361 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3421 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     3481 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3541 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3601 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3661 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3721 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3781 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3841 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3901 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4021 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4081 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4141 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4201 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4261 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4321 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4381 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4441 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4501 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4561 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4621 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4681 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4741 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4801 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4861 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4921 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4981 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5041 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5101 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5161 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5221 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5281 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5341 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5401 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5461 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5521 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5581 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5641 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5701 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5761 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5821 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5881 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5941 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6001 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6061 aagttgggta acgccagggt tttcccagtc acgacgtt
//
