LOCUS       sequence-270117-        6368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G51810 M2_pECIA14 should be used with bait vector 
            AT1G51810 M2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6368)
  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 6368)
  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: AT1G51810 M2_pECIA14
FEATURES             Location/Qualifiers
     source          1..6368
                     /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             1664..1687
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3362..3385
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3401..3418
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3441..3458)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3491..3625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3569..3588
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3769..3785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3769..3785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3782..3804)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3793..3809
                     /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(3817..3823,3824..3841,3842..3847))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3862..3883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4000..4017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4171..4759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4251..4270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4930..5721,5722..5790))
                     /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     5553..5572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5791..5895)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5963..5981
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6019..6041)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6141..6160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6354..6368,1..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 atgtggatca cggtgaatac
      541 tgataacact atcaaggaga tcctccacgt atcgaaatca aacactttgc aagtgtgtct
      601 tgttaagaca ggtacaagta taccttatat aaatacattg gaactacgac cattggccga
      661 cgatatatac accaacgaaa gtggctccct caactatctt tttcgggttt attatagcaa
      721 tttaaagggc tatatagagt accccgatga tgtccacgat cgcatatgga aacaaatcct
      781 accttaccag gattggcaga ttttaactac gaatctccaa ataaacgttt ctaatgatta
      841 tgatctaccc caacgtgtaa tgaaaacagc tgtaacaccc attaaagcta gcacaacgac
      901 gatggaattt ccctggaact tagagcctcc aacttcacag ttttacttat tccttcactt
      961 tgcagagctt caaagtctac aagccaacga gacgagggaa ttcaatgtgg tgttgaacgg
     1021 aaatgttaca tttaaatctt atagtcctaa gtttttagaa atgcaaacag tatatagcac
     1081 agcaccaaag caatgcgatg gagggaaatg cttgttgcag ttagtgaaaa cgtcaaggtc
     1141 cactttgccg cctctaatta atgctatgga ggcttacact gtgcttgatt tcccacagat
     1201 agaaacaaat gtagatgaag tgattgctat caagaatata caatctactt atggattgag
     1261 taaaacaacc tggcaaggag atccatgtgt acccaaaaag ttcttgtggg atggtttaaa
     1321 ctgcaacaac tcggatgatt ctacgccacc aattatcact tccttaaact tgtcttcaag
     1381 tggattgact ggaattatcg tgctgaccat tcagaatctc gccaatttac aagaactgga
     1441 cttgtcaaat aacaacttgt ctggaggtgt tcctgaattt ctagcggata tgaagtcgct
     1501 cttggtcata aacttaagtg ggaacaatct cagtggtgta gttcctcaaa agcttataga
     1561 gaagaaaatg ttgaaattga acattgaagg caatccgaag cttaattgca cagtggagtc
     1621 atgtgtaaac aaagatgaag agggtggacg acagataaag agcctggagg tgctgttcca
     1681 gggaccaagc ggcaatcccg gcggcgatct ggccccccag atgctgcgcg agctgcagga
     1741 gaccaatgcc gccctgcagg acgtgcgcga gctgctgcgc caccgcgtga aggagatcac
     1801 cttcctgaag aacaccgtga tggagtgcga cgcctgcggc atgcagccct ctagaggctc
     1861 cggctccatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     1921 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     1981 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     2041 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     2101 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     2161 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     2221 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     2281 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     2341 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     2401 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     2461 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     2521 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     2581 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     2641 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     2701 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     2761 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     2821 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     2881 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     2941 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     3001 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     3061 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     3121 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     3181 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     3241 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     3301 ccccgccggc accaccgacg ccgcgcaccc gggttctaga ggaagcacct ccggcgataa
     3361 cgattacaag gatgatgatg ataaggccag cggcaccggt catcatcacc atcaccattg
     3421 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     3481 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3541 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3601 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3661 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3721 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3781 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3841 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3901 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3961 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4021 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4081 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4141 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4201 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4261 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4321 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4381 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4441 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4501 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4561 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4621 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4681 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4741 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4801 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4861 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4921 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4981 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5041 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5101 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5161 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5221 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5281 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5341 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5401 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5461 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5521 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5581 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5641 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5701 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5761 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5821 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5881 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5941 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6001 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6061 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6121 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6181 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6241 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6301 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6361 acgacgtt
//
