LOCUS       sequence-274539-        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G56140 X032_pECIA14 should be used with bait vector 
            AT1G56140 X032_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  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 7028)
  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: AT1G56140 X032_pECIA14
FEATURES             Location/Qualifiers
     source          1..7028
                     /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             2324..2347
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4022..4045
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4061..4078
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4101..4118)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4151..4285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4175..4194)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4229..4248
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4429..4445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4429..4445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4442..4464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4453..4469
                     /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(4477..4483,4484..4501,4502..4507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4522..4543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4660..4677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4831..5419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4911..4930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5590..6381,6382..6450))
                     /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     6213..6232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6451..6555)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6623..6641
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6679..6701)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6801..6820
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7014..7028,1..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 acaggagcca ccacacatcc
      541 cgacgaagcg ctagctttga actcaatttt cgcggcttgg aggattcggg cgccgaggga
      601 atggaacatc agcggcgaac tttgctccgg cgccgctatc gacgccagtg ttctcgactc
      661 aaaccctgcc tacaatcctc tcatcaaatg cgactgcagt ttcgaaaact ccacaatctg
      721 ccgcattacc aacatcaagg tttatgcgat ggaagttgta ggatctatac ctcaacaact
      781 ctggaccttg gaatacctga caaatctgaa cttgggtcaa aatgttctca ctggctcact
      841 tcctcctgca cttggaaatt tgactcgaat gcgatggatg acttttggga tcaatgcgtt
      901 gtctggccct attcctaaag aaatcggttt gcttacagat ttaagattac ttagtattag
      961 ttcaaataac ttttcgggtt ctataccaga tgagattggg agatgtacaa aactacaaca
     1021 gatatacata gatagttcgg gactcagcgg gggattacct gtatcatttg ctaatcttgt
     1081 ggagctggaa caagcttgga ttgcggatat ggaacttaca ggtcagatac cagactttat
     1141 aggagattgg accaaactta ctaccttgag aattcttgga actggtttga gtggtccgat
     1201 accggcgtcg ttttcaaact taacttcttt gacagagctg aggcttggtg atatatccaa
     1261 tggaaattct tctcttgaat tcatcaaaga catgaaatct ctaagtatat tagtattgag
     1321 gaacaataat ctcactggga caataccgtc taatattgga gaatattcaa gtttgcgaca
     1381 acttgatttg agcttcaaca aactacatgg aacaattccg gcttcacttt tcaacttaag
     1441 acagcttact cacttgtttc tggggaacaa cacgttgaat ggctcattgc ccactcaaaa
     1501 ggggcagtct ctgagcaatg tagatgtgtc atacaatgat ttgtctggaa gtcttccgtc
     1561 atgggtcagt ttaccaaact tgaatctcaa cttagttgct aacaacttca cattggaagg
     1621 tcttgacaac agggttttat caggactgaa ctgcctgcag aagaacttcc catgcaatcg
     1681 agggaaagga atatattctg acttctcaat caactgcgga ggcccagaga taagatctgt
     1741 aactgaagca gtatttgaga gggaggacga ggatcttgga ccagcttcat ttgtcgtgag
     1801 tgctggtcag agatgggcag ccagtagtgt aggacttttt gccggaagta gcaacaatat
     1861 atacatatct acttcacaat cacaatttgt caacactttg gactcagagc tatttcagtc
     1921 agcaagactt tctgcatctt ccctaaggta ttatgggttg ggtctagaaa atggaggcta
     1981 taccgtcaca cttcagtttg ctgaaataca aattctaggt tctacttcca acacttggag
     2041 aggtttagga agacgacgtt ttgacattta tgtccaggga agacttgttg aaaaggactt
     2101 cgatgtacgc agaacagctg gtgactccac tgttcgagca gttcagagag aatataaagc
     2161 aaatgtatca caaaatcatc tcgaaattca tcttttctgg gctggaaaag gaacatgctg
     2221 tattcctatc caaggggctt atgggccatt aatatcggcc gtcggtgcaa caccagattt
     2281 cacaccaact gtgggtaata ggccaccatc aaagggaaag agcctggagg tgctgttcca
     2341 gggaccaagc ggcaatcccg gcggcgatct ggccccccag atgctgcgcg agctgcagga
     2401 gaccaatgcc gccctgcagg acgtgcgcga gctgctgcgc caccgcgtga aggagatcac
     2461 cttcctgaag aacaccgtga tggagtgcga cgcctgcggc atgcagccct ctagaggctc
     2521 cggctccatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     2581 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     2641 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     2701 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     2761 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     2821 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     2881 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     2941 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     3001 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     3061 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     3121 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     3181 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     3241 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     3301 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     3361 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     3421 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     3481 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     3541 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     3601 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     3661 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     3721 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     3781 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     3841 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     3901 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     3961 ccccgccggc accaccgacg ccgcgcaccc gggttctaga ggaagcacct ccggcgataa
     4021 cgattacaag gatgatgatg ataaggccag cggcaccggt catcatcacc atcaccattg
     4081 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     4141 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4201 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4261 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4321 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     4381 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     4441 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4501 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4561 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4621 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4681 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4741 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4801 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4861 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4921 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4981 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5041 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5101 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5161 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5221 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5281 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5341 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5401 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5461 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5581 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5641 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5701 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5761 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5821 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5881 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5941 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6001 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6061 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6121 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6181 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6241 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6301 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6361 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6421 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6481 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6541 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6601 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6661 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6721 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6781 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6841 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6901 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6961 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7021 acgacgtt
//
