LOCUS       sequence-238705-        4961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT4G23740 J11_pECIA2 should be used with prey vector 
            AT4G23740 J11_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4961)
  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 4961)
  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: AT4G23740 J11_pECIA2
FEATURES             Location/Qualifiers
     source          1..4961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1659..1682
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2388..2429
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2439..2456
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2479..2496)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2529..2663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2553..2572)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2607..2626
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2807..2823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2807..2823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2820..2842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2831..2847
                     /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(2855..2861,2862..2879,2880..2885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2900..2921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3038..3055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3209..3797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3289..3308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3968..4759,4760..4828))
                     /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     4591..4610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4829..4933)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggtcaga tccattagaa gacaaacgcg cattgcttga gtttcttacc attatgcaac
     1021 caactcgttc actcaactgg aacgagactt ctcaagtctg caacatctgg accggagtta
     1081 cttgtaatca agacggatct cgaatcatag ccgttagatt accaggcgtt ggactcaatg
     1141 gtcagattcc tccaaacact ataagcagac tctcagctct cagagttctt agtctcagat
     1201 ccaatctaat ctccggcgag tttcctaagg attttgttga gttgaaagat ttggcctttc
     1261 tttatcttca ggataacaac ttgtctggtc ctttgccatt ggatttctct gtttggaaga
     1321 atctcactag tgttaatctc tcaaacaacg ggttcaacgg gacaatcccg agttcgttgt
     1381 ctcgtcttaa gcggattcag tcgttaaatc ttgctaacaa tacgctttct ggtgatattc
     1441 ctgatctgag tgtgctctct agtctgcagc acattgactt gtccaacaat tatgatcttg
     1501 ctggaccgat accagattgg cttcgaaggt tcccgttctc gtcttataca ggcattgata
     1561 ttattcctcc tggtggtaac tacaccctcg ttacgccgcc accacctagc gagcagactc
     1621 atcagaaacc gtcgaaagct cgcttcttgg gactatccct ggaggtgctg ttccagggac
     1681 cagggagcga caaaactcac acatgcccac cgtgcccagc acctgaactc ctggggggac
     1741 cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc cggacccctg
     1801 aggtcacatg cgtggtggtg gacgtgagcc acgaagaccc tgaggtcaag ttcaactggt
     1861 acgtggacgg cgtggaggtg cataatgcca agacaaagcc gcgggaggag cagtacaaca
     1921 gcacgtaccg tgtggtcagc gtcctcaccg tcctgcacca ggactggctg aatggcaagg
     1981 agtacaagtg caaggtctcc aacaaagccc tcccagcccc catcgagaaa accatctcca
     2041 aagccaaagg gcagccccga gaaccacagg tgtacaccct gcccccatcc cgggaggaga
     2101 tgaccaagaa ccaggtcagc ctgacctgcc tggtcaaagg cttctatccc agcgacatcg
     2161 ccgtggagtg ggagagcaat gggcagccgg agaacaacta caagaccacg cctcccgtgc
     2221 tggactccga cggctccttc ttcctctaca gcaagctcac cgtggacaag agcaggtggc
     2281 agcaggggaa cgtcttctca tgctccgtga tgcatgaggg tctgcacaac cactacacgc
     2341 agaagagcct ctccctgtct ccgggtaaat ctagagggcc cttcgaaggt aagcctatcc
     2401 ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag
     2461 tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct
     2521 cgatcgagga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2581 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2641 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2701 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     2761 attatgatca gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc
     2821 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2881 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2941 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3961 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4021 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4081 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4141 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4201 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4261 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4321 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4381 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4441 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4501 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4561 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4621 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4681 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4741 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4801 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4861 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4921 taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
