LOCUS       sequence-276514-        6461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G16250 K9_pECIA14 should be used with bait vector 
            AT2G16250 K9_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6461)
  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 6461)
  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: AT2G16250 K9_pECIA14
FEATURES             Location/Qualifiers
     source          1..6461
                     /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             1757..1780
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3455..3478
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3494..3511
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3534..3551)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3584..3718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3608..3627)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3662..3681
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3862..3878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3862..3878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3875..3897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3886..3902
                     /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(3910..3916,3917..3934,3935..3940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3955..3976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4093..4110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4264..4852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4344..4363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5023..5814,5815..5883))
                     /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     5646..5665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5884..5988)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6056..6074
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6112..6134)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6234..6253
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6447..6461,1..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 agtcgattca gtgagaaact
      541 cattttgctt aatctcaggt cttctttagg gttaagagga acagattggc caattaaagg
      601 tgacccttgt gttgattgga gaggtataca atgtgagaat ggtagtatca ttgggatcaa
      661 tatctctggg tttaggagaa caagaattgg taagctaaac ccacaattct ctgttgatcc
      721 tcttcgtaat ctaactcgtt tgtcgtattt taacgcgtcc gggttagcgt taccggggac
      781 tatacctgag tggtttggtg ttagtttgtt ggcattggaa gtattggatc ttagttcgtg
      841 ttctgttaac ggtgttgttc cttttactct tggtaatctc actagtttaa gaactttgaa
      901 tctgtctcag aacagtctta ctagtttagt tcctagcagt ttaggacagt tgttgaatct
      961 atcacagctt gatctctcta ggaactcatt cactggtgtt cttcctcaat cttttagctc
     1021 tctgaagaat ctgttgactc ttgatgtttc ttccaattac cttactggac ctatccctcc
     1081 tggtttagga gcgttatcaa agcttatcca tctgaacttt tcaagcaata gtttctcgtc
     1141 tccgattcct tcagagcttg gtgatcttgt caatctggtt gactttgatc tcagcatcaa
     1201 ttcattgtct ggttcagttc ctcaggagct aagaaagtta agtaagttgc agctaatggc
     1261 gattggcgat aaccttttga gtggtacttt accagtagat ttgtttagtg ctgagagcca
     1321 actgcaaact ttagttttga gggaaaatgg tttctctggt agtctacctg atgtgtgctg
     1381 gtcgttgcca aaattgcgga ttcttgacat tgctaagaac aacttcacag gactgctgcc
     1441 atattctagt tatgattcag accagatagc tgaaatggtt gatatctctt cgaacacatt
     1501 ctatggagag ctcacaccga ttcttagacg gttcaggata atggacctct ctggtaatta
     1561 tttcgaaggt aagcttcctg attatgtgac cggggaaaat gtgtctgtta caagcaattg
     1621 ccttcgaaat gagaggagac agaagccgtc agcaatatgt gcagcattct ataagtctag
     1681 aggattagat tttgatgatt tcggacgtcc taatttgaca caaccgacat cgaaaaatgc
     1741 ttcttcaggg ataagcctgg aggtgctgtt ccagggacca agcggcaatc ccggcggcga
     1801 tctggccccc cagatgctgc gcgagctgca ggagaccaat gccgccctgc aggacgtgcg
     1861 cgagctgctg cgccaccgcg tgaaggagat caccttcctg aagaacaccg tgatggagtg
     1921 cgacgcctgc ggcatgcagc cctctagagg ctccggctcc atcatcccag ttgaggagga
     1981 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     2041 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     2101 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     2161 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     2221 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     2281 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     2341 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     2401 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     2461 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     2521 tacgcagctc atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat
     2581 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     2641 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     2701 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     2761 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     2821 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     2881 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     2941 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     3001 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     3061 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     3121 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     3181 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     3241 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     3301 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     3361 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     3421 cccgggttct agaggaagca cctccggcga taacgattac aaggatgatg atgataaggc
     3481 cagcggcacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     3541 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     3601 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3661 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3721 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     3781 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga cctgcaggca
     3841 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6001 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     6061 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     6121 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6181 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6241 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6301 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6361 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6421 attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
