LOCUS       sequence-270291-        6026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G50230 J8_pECIA14 should be used with bait vector 
            AT3G50230 J8_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6026)
  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 6026)
  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: AT3G50230 J8_pECIA14
FEATURES             Location/Qualifiers
     source          1..6026
                     /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             1322..1345
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3020..3043
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3059..3076
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3099..3116)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3149..3283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3173..3192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3227..3246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3427..3443)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3427..3443)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3440..3462)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3451..3467
                     /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(3475..3481,3482..3499,3500..3505))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3520..3541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3658..3675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3829..4417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3909..3928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4588..5379,5380..5448))
                     /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     5211..5230
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5449..5553)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5621..5639
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5677..5699)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5799..5818
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6012..6026,1..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 tctctcccgg cgacgaacta
      541 cttcgactcc ttcttaccat ccgacgccgt cgcacttctc tccttcaaat ccaccgctga
      601 tctcgacaac aagcttctat actctctcac cgagccttac gactactgtc aatggcgcgg
      661 cgtcgattgc tctcaagatc gcgtcgttcg tcttatcctc gacggtgttg gtctccgggg
      721 aagtttctca ccggagactc ttagtcgtct tgatcagctt cgtgttctta gccttgagaa
      781 taactcaatc tcgggttcga ttcctgatct atctcctctc gtcaatctca aaaccctaac
      841 cctaagcaag aatggtttct ccggtactct ttcttcttca attctctctc ttcgtcggtt
      901 aacagaactt gatctatcgt ttaacaactt ctccggtgag attccgtcgg ggatcaatgc
      961 tctgagtcga ctcagttctt taaatctcga gtttaatcga cttaacggaa ctctcccgcc
     1021 gttaaatctt tcttctctga tttcatttaa cgtttccagt aacaacctca ccggtttagt
     1081 tccgttaaca aagactctgt tacgattcaa cgcgtcttcg ttttcgtcaa accctggact
     1141 ctgcggtgag atcattaacc gatcttgcgg tttacattct tcttctccgt tctttggttc
     1201 acctaaacca aacaccacgt catcaacatc ttctgcttct tcgtctgaag caccggttat
     1261 tcaaagtgaa cagaacggtg aagcagcgat gattgttcca ccggtggtta aaaaggttaa
     1321 actggaggtg ctgttccagg gaccaagcgg caatcccggc ggcgatctgg ccccccagat
     1381 gctgcgcgag ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc tgctgcgcca
     1441 ccgcgtgaag gagatcacct tcctgaagaa caccgtgatg gagtgcgacg cctgcggcat
     1501 gcagccctct agaggctccg gctccatcat cccagttgag gaggagaacc cggacttctg
     1561 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     1621 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     1681 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     1741 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     1801 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     1861 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     1921 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     1981 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     2041 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     2101 caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac
     2161 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     2221 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     2281 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     2341 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     2401 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     2461 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     2521 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     2581 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     2641 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     2701 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     2761 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     2821 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     2881 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     2941 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gttctagagg
     3001 aagcacctcc ggcgataacg attacaagga tgatgatgat aaggccagcg gcaccggtca
     3061 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     3121 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3181 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3241 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3301 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3361 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     3421 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3481 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3541 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3601 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3661 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3721 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3781 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3841 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3901 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3961 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4021 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4081 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4141 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4201 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4261 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4321 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4381 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4441 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4501 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4561 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4621 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4681 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4741 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4801 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4861 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4921 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4981 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5041 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5101 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5161 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5221 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5281 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5341 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5401 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5461 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5521 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5581 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5641 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5701 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5761 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5821 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5881 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5941 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6001 gccagggttt tcccagtcac gacgtt
//
