LOCUS       sequence-296637-        5540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G46370 N8_pECIA2 should be used with prey vector 
            AT3G46370 N8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5540)
  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 5540)
  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: AT3G46370 N8_pECIA2
FEATURES             Location/Qualifiers
     source          1..5540
                     /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             1793..1816
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2522..2563
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2573..2590
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2613..2630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2663..2797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2687..2706)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2741..2760
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2941..2957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2941..2957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2954..2976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2965..2981
                     /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(2989..2995,2996..3013,3014..3019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3034..3055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3172..3189)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3343..3931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3423..3442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4102..4893,4894..4962))
                     /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     4725..4744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4963..5067)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5135..5153
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5191..5213)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5313..5332
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5526..5540,1..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 atgcgaaact gttacaatct
      541 tagtgtgcat aaagaaacaa agtatttgat tagggttaca agtaactatg gaaattacga
      601 tggtcgtaac gaacctccta gatttgattt gtacttaggc cctaacttct gggtaacaat
      661 agatttgggg aagcatgtaa atggtgatac atggaaggag attattcata tcccaaagtc
      721 aaactctcta gatgtgtgtc ttatcaaaac aggtacaaca acgccaatta tatcgacctt
      781 ggaactaagg tctctaccaa agtatagtta caacgcaata tcaggttctt tgaagtcgac
      841 actgagggca ttcctaagcg aatcaacaga agttataagg tacccaaatg atttctacga
      901 tcggatgtgg gttccacatt ttgagacgga atggaagcaa atttctacca atctcaaagt
      961 gaacagctcc aatggttatc ttctgccaca ggatgtgctc atgaccgccg caatacctgt
     1021 gaatactagt gcacgattga gttttaccga gaatcttgag ttccctcatg atgaacttta
     1081 cctgtacttc cacttctctg aggtccaagt cttacaggcc aaccagagta gagagttcag
     1141 cattttgtgg aacggaatgg ttatttaccc agattttatc cctgactatc ttggtgccgc
     1201 cactgtgtac aacccgtcac catctctttg tgaagtaggg aaatgcttat tagagcttga
     1261 aagaactcag aaatcaactc tcccgcctct ccttaacgcc attgaagttt tcacagtgat
     1321 gaattttcca cagtctgaga caaacgatga tgatgtaatt gctataacaa agatcaaaga
     1381 cacacacaga ttgaatagaa cctcctggca aggagatcca tgcgtccctc aactgttttc
     1441 gtgggccggt ctaagttgca ttgacacgaa tgtatctacc ccaccaagaa tcatttccct
     1501 aaacttgtct tcaagtgggt taacaggaaa catagcaact ggaattcaaa atctgaccaa
     1561 actgcaaaag ctggacttgt caaataacaa tttgacagga gtagtgcccg aatttctagc
     1621 taacatgaaa tcgttgctgt tcatagactt gaggaagaac aaactgaatg gttcaatacc
     1681 aaagactcta cttgatagaa aaaagaaagg actacaatta tttgttgatg gcgatgatga
     1741 taagggcgat gataataagt gtttatccgg ttcatgtgtc ccaaaaatga aactggaggt
     1801 gctgttccag ggaccaggga gcgacaaaac tcacacatgc ccaccgtgcc cagcacctga
     1861 actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat
     1921 ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt
     1981 caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga
     2041 ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg
     2101 gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga
     2161 gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc
     2221 atcccgggag gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta
     2281 tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac
     2341 cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga
     2401 caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg agggtctgca
     2461 caaccactac acgcagaaga gcctctccct gtctccgggt aaatctagag ggcccttcga
     2521 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
     2581 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     2641 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     2701 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2761 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2821 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2881 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     2941 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3001 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3061 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3121 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3181 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3241 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3301 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3361 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3421 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3481 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3541 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3601 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3661 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3721 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3781 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3841 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3901 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3961 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4021 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4081 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4141 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4201 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4261 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4321 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4381 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4441 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4501 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4561 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4621 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4681 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4741 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4801 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4861 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4921 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4981 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5041 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5101 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5161 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5221 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5281 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5341 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5401 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5461 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5521 gttttcccag tcacgacgtt
//
