LOCUS       sequence-237787-        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G66830 X005_pECIA14 should be used with bait vector 
            AT1G66830 X005_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  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 6071)
  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: AT1G66830 X005_pECIA14
FEATURES             Location/Qualifiers
     source          1..6071
                     /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             1812..1835
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3510..3533
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3549..3566
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3589..3606)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3639..3773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3717..3736
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3917..3933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3917..3933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3930..3952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3941..3957
                     /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(3965..3971,3972..3989,3990..3995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4010..4031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4148..4165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4319..4907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4399..4418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5078..5869,5870..5938))
                     /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     5701..5720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5939..6043)
                     /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 tcgggtctct aaacgatcaa ggtcttgctc ttctttcctt taaacaatcc atacaaaacc
     1021 agagtgactc tgtttttacc aattggaact cttcagattc aaacccttgt tcttggcaag
     1081 gtgttacatg taattatgat atgagagttg tctcgattcg tcttcctaat aaacgtcttt
     1141 cgggttctct tgatccatcc attggaagtc tcttgtccct tcgtcacatt aacctaagag
     1201 acaatgactt tcaagggaag ttacctgtgg aactctttgg acttaaaggg cttcaaagct
     1261 tggtgctttc tggaaattct ttttccgggt ttgtcccaga agagattggt agcttgaaga
     1321 gtctaatgac tctggatctt tcggagaatt ctttcaacgg ttcgatttct ttgtcgctta
     1381 ttccatgcaa gaagctgaaa acgcttgttc ttagcaagaa cagtttctct ggagatcttc
     1441 caacagggtt aggttctaac ttggttcatc ttcggacact gaatctttct ttcaatcgct
     1501 tgactggaac gatccccgaa gatgtaggaa gtctggagaa tctgaaaggt acacttgatc
     1561 tttctcataa cttcttctca ggtatgattc caacaagtct tggaaatctc cctgagcttc
     1621 tctatgtcga tctttcttac aataatctca gtggtccgat accaaaattc aacgttctgt
     1681 tgaatgccgg tccaaatgct ttccaaggga accctttcct ctgtggatta ccgataaaga
     1741 tttcttgcag cacaagaaac acacaagtag ttccttctca actatataca agaagagcca
     1801 accatcattc tctggaggtg ctgttccagg gaccaagcgg caatcccggc ggcgatctgg
     1861 ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc
     1921 tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa caccgtgatg gagtgcgacg
     1981 cctgcggcat gcagccctct agaggctccg gctccatcat cccagttgag gaggagaacc
     2041 cggacttctg gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg
     2101 cacagacagc cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg
     2161 tgacagctgc caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc
     2221 tggccatgga ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg
     2281 tgccagacag tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga
     2341 ccattggctt gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg
     2401 tcatctccgt gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca
     2461 cacgagtgca gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt
     2521 actcggacgc cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc
     2581 agctcatctc caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc
     2641 gcatgggaac cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg
     2701 acgggaagaa tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga
     2761 accgcactga gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct
     2821 ttgagcctgg agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga
     2881 tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct
     2941 tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga
     3001 ctgagacgat catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg
     3061 acacgctgag cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc
     3121 tgcgagggag ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca
     3181 cggtcctcct atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg
     3241 ttaccgagag cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg
     3301 aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg
     3361 ttcacggcgt gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg
     3421 agccctacac cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg
     3481 gttctagagg aagcacctcc ggcgataacg attacaagga tgatgatgat aaggccagcg
     3541 gcaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     3601 cttctaaggc ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca
     3661 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3721 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3781 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     3841 agtaaaacct ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa
     3901 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3961 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4021 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4081 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4141 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4741 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4981 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5041 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5101 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5161 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5221 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5281 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5341 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5401 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5461 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5521 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5581 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5641 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5701 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5761 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5821 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5881 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5941 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6001 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6061 tgccacctga c
//
