LOCUS       sequence-274562-        6818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G49760 X026_pECIA14 should be used with bait vector 
            AT5G49760 X026_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6818)
  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 6818)
  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: AT5G49760 X026_pECIA14
FEATURES             Location/Qualifiers
     source          1..6818
                     /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             2114..2137
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3812..3835
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3851..3868
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3891..3908)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3941..4075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3965..3984)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4019..4038
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4219..4235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4219..4235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4232..4254)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4243..4259
                     /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(4267..4273,4274..4291,4292..4297))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4312..4333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4450..4467)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4621..5209)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4701..4720)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5380..6171,6172..6240))
                     /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     6003..6022
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6241..6345)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6413..6431
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6469..6491)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6591..6610
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6804..6818,1..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 acgaatggtt tagacgcttc
      541 tgctttaaat gctctgaaga gtgaatggac cactcctcct gatggttggg aaggctctga
      601 tccttgtgga accaattggg ttggaattac atgtcaaaat gaccgcgtcg tttcaatatc
      661 actaggtaac cttgacttgg aaggaaagct tcctgccgat atatcgttct tgtccgagtt
      721 gcggatcttg gatttgtcat acaatcctaa attgtccgga ccgcttccac caaacatcgg
      781 taaccttggg aagttaagga acttgatcct tgtgggatgt agtttcagtg gtcaaatccc
      841 tgagtccatt ggaactctaa aagaacttat atatctctcc ctaaatttaa ataaatttag
      901 tggaacaatt ccgccttcaa ttggcctgtt atcaaaacta tattggtttg atatagctga
      961 caatcagatt gaaggagagc ttccagtttc taatgggact tctgcacctg gacttgacat
     1021 gcttcttcaa actaagcatt ttcattttgg aaaaaacaag ctttcaggca atatcccaaa
     1081 agaacttttc agctcaaaca tgagtttaat acatgtcctc ttcgatggaa accaattcac
     1141 gggcgaaatt ccagagaccc tcagtctcgt taaaacgttg acggtgttac gccttgatag
     1201 gaacaaactc attggagata ttccttcata tcttaacaat ctcacaaatc ttaacgaact
     1261 gtacttggcc aacaacagat ttactggtac tcttccaaat ttaaccagct tgaccagtct
     1321 ctacacattg gatgtgagca ataacacttt ggatttctca cccattccat catggatctc
     1381 ttcattaccc tccctatcaa cattaaggat ggaagggatc caacttaatg gtccaatacc
     1441 aatctcattt ttcagccctc ctcagttgca gactgttatc ttaaagcgaa atagtatagt
     1501 ggaaagctta gactttggta cagacgttag cagccagttg gagtttgtgg atttacaata
     1561 caatgaaata actgattata aaccatcagc taataaagtc ctccaagtaa tattggcaaa
     1621 taatccagtg tgtctagagg cgggaaacgg gccgagttac tgttcagcaa tccaacacaa
     1681 tacctcattt tctacactcc caacaaactg ttctccatgc gaaccgggca tggaagcgag
     1741 tcccacgtgc cgctgtgcgt atccattcat gggaacactc tacttccggt ctccttcttt
     1801 ctcagggttg ttcaactcca ccaacttctc aattctacag aaggcaatcg cggatttctt
     1861 taagaagttc aattatcctg tagactccgt tggtgttcgg aacataagag agaatccaac
     1921 tgatcatcag cttctaatag atctattggt ctttccattg ggaagagaga gttttaatca
     1981 gacaggaatg tcacttgttg gttttgcatt tagcaaccag acttataagc ctcctccgat
     2041 atttggccct tatattttca aagctgattt gtacaaacag ttctctgatg tagaagtttc
     2101 atcaaagtca tcactggagg tgctgttcca gggaccaagc ggcaatcccg gcggcgatct
     2161 ggccccccag atgctgcgcg agctgcagga gaccaatgcc gccctgcagg acgtgcgcga
     2221 gctgctgcgc caccgcgtga aggagatcac cttcctgaag aacaccgtga tggagtgcga
     2281 cgcctgcggc atgcagccct ctagaggctc cggctccatc atcccagttg aggaggagaa
     2341 cccggacttc tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc
     2401 tgcacagaca gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac
     2461 ggtgacagct gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc
     2521 cctggccatg gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca
     2581 tgtgccagac agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca
     2641 gaccattggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga
     2701 ggtcatctcc gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac
     2761 cacacgagtg cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg
     2821 gtactcggac gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac
     2881 gcagctcatc tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt
     2941 tcgcatggga accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct
     3001 ggacgggaag aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg
     3061 gaaccgcact gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct
     3121 ctttgagcct ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct
     3181 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct
     3241 cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact
     3301 gactgagacg atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga
     3361 ggacacgctg agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc
     3421 cctgcgaggg agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta
     3481 cacggtcctc ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga
     3541 tgttaccgag agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga
     3601 cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct
     3661 ggttcacggc gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct
     3721 ggagccctac accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc
     3781 gggttctaga ggaagcacct ccggcgataa cgattacaag gatgatgatg ataaggccag
     3841 cggcaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     3901 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     3961 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     4021 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     4081 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     4141 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     4201 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5041 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6421 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6481 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6541 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6601 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6661 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6781 aagttgggta acgccagggt tttcccagtc acgacgtt
//
