LOCUS       sequence-298761-        5636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT2G01820 X036_pECIA2 should be used with prey vector 
            AT2G01820 X036_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5636)
  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 5636)
  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: AT2G01820 X036_pECIA2
FEATURES             Location/Qualifiers
     source          1..5636
                     /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             1889..1912
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2618..2659
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2669..2686
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2709..2726)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2759..2893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2837..2856
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3037..3053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3037..3053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3050..3072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3061..3077
                     /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(3085..3091,3092..3109,3110..3115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3130..3151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3268..3285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3439..4027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3519..3538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4198..4989,4990..5058))
                     /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     4821..4840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5059..5163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5231..5249
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5287..5309)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5409..5428
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5622..5636,1..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 tctcaaaccg gtctcgatga
      541 ttcgactatg caatctctca aatcaagtct gaatcttaca tcagacgtcg attggtctaa
      601 ccctaaccct tgtaaatggc aatctgttca atgcgatgga agtaatcgtg ttactaagat
      661 tcagcttaaa caaaaaggga ttcgtggaac tcttcctact aatcttcaga gtctctctga
      721 attggttatt ctcgagcttt ttctcaatcg tatctctggt cctattcctg atctctctgg
      781 cttatctcgt ttacagactc tgaatctaca cgacaatctt tttacctctg taccgaagaa
      841 tctcttctct ggtatgagtt ctcttcaaga aatgtatctc gagaacaatc cgtttgatcc
      901 ttgggtaatt ccagacaccg ttaaagaagc tacgtctctg cagaatctaa ctctctccaa
      961 ttgcagcatt attggaaaga tcccagattt cttcggttct cagtcacttc cgagcttgac
     1021 gaatctgaaa ctatctcaga atggtttaga aggagagtta ccgatgagtt ttgctggtac
     1081 ttcgattcag tctctgttct tgaatggtca gaagcttaat ggatcgatct cggtattggg
     1141 gaacatgact tcgctcgttg aggtttctct tcaaggaaat cagttctcag gtccaattcc
     1201 tgatctctct ggtttggtat ctctacgtgt gtttaatgtt agagagaatc agctcactgg
     1261 tgttgttcca cagtctttgg ttagtttgag ttcgcttact actgtgaatt taactaataa
     1321 ttatcttcaa ggaccgactc cgttgtttgg caaatccgtt ggtgttgata tagtcaacaa
     1381 catgaatagc ttttgtacta atgtagctgg tgaggcttgt gatcctcgtg tggatacttt
     1441 ggtatctgta gctgaatcat ttggatatcc agtgaagctt gcggaaagtt ggaaaggaaa
     1501 caatccttgt gttaactggg ttgggattac ttgttctgga ggtaacatta cggttgttaa
     1561 tatgaggaaa caggatctct cgggtacgat ttctccgagt ttagctaagc ttacttcatt
     1621 ggagactatc aatcttgctg ataataaact ctccgggcat atacctgatg agcttaccac
     1681 attgtctaag cttaggttac ttgatgtgtc taacaacgat ttttatggaa taccgccaaa
     1741 gtttcgggat actgtgactt tggtgactga agggaacgct aacatgggga agaatggacc
     1801 taacaaaacg agtgatgctc ctggagcttc accaggaagt aaaccttcag gaggaagtga
     1861 tggcagtgaa acttctaaga aatcaagcct ggaggtgctg ttccagggac cagggagcga
     1921 caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac cgtcagtctt
     1981 cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg aggtcacatg
     2041 cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt acgtggacgg
     2101 cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca gcacgtaccg
     2161 tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg agtacaagtg
     2221 caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca aagccaaagg
     2281 gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga tgaccaagaa
     2341 ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg ccgtggagtg
     2401 ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc tggactccga
     2461 cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc agcaggggaa
     2521 cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc agaagagcct
     2581 ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc ctaaccctct
     2641 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     2701 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2761 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2821 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2881 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2941 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3001 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3061 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3121 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3181 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3241 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3601 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3841 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4501 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4801 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5161 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5221 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5281 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5341 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5401 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5461 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5521 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5581 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
