LOCUS       sequence-270344-        6107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G25320 X002_pECIA14 should be used with bait vector 
            AT1G25320 X002_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6107)
  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 6107)
  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: AT1G25320 X002_pECIA14
FEATURES             Location/Qualifiers
     source          1..6107
                     /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             1403..1426
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3101..3124
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3140..3157
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3180..3197)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3230..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3254..3273)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3308..3327
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3508..3524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3508..3524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3521..3543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3532..3548
                     /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(3556..3562,3563..3580,3581..3586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3601..3622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3739..3756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3910..4498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3990..4009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4669..5460,5461..5529))
                     /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     5292..5311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5530..5634)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5702..5720
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5758..5780)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5880..5899
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6093..6107,1..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 aatgctctaa acgacgaagg
      541 gtttgctctt ttaactctga agcagtctat ttcaaaagat ccagatggtt ctttaagcaa
      601 ttggaactca gagaatcaaa acccttgttc ttggaatgga gtcacttgtg atgataacaa
      661 ggttgttgtc tctcttagca tcccaaagaa gaaactttta ggttatcttc cttcttctct
      721 tggtttactc tctaatctcc gtcatttgaa tttgagaagc aatgagctta gtgggaactt
      781 acctgttgag ctttttaagg ctcaaggtct tcaaagtttg gttctttatg ggaatttctt
      841 gtctgggtcc attccaaatg agattggtga cttaaagttt ctgcaaattt tggatctttc
      901 tcgtaattct cttaatgggt ctattccaga atcggttttg aagtgtaata ggcttaggag
      961 ctttgatttg agtcagaata atctcactgg ttctgttcct agtggatttg gtcagtcttt
     1021 ggcttctttg cagaagcttg atctttcttc taacaatctt attggtcttg ttcctgatga
     1081 tttggggaat ttgactagat tgcaaggaac tcttgatttg tctcataatt cgtttagtgg
     1141 ctcgattcca gctagtttgg ggaatttgcc tgagaaagtt tatgtcaatc tagcttacaa
     1201 caacttgagt ggaccaatcc cacaaaccgg tgctttggtg aacagaggac caactgcatt
     1261 cttggggaat ccaagactct gtggccctcc attgaaggat ccttgtttgc cagatacgga
     1321 tagttcttca acttctcacc cttttgtacc ggataacaac gagcaaggtg gaggagggtc
     1381 aaagaaagga gaaggtttaa gtctggaggt gctgttccag ggaccaagcg gcaatcccgg
     1441 cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg ccctgcagga
     1501 cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat
     1561 ggagtgcgac gcctgcggca tgcagccctc tagaggctcc ggctccatca tcccagttga
     1621 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     1681 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     1741 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     1801 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     1861 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     1921 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     1981 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     2041 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     2101 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     2161 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     2221 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     2281 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     2341 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     2401 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     2461 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     2521 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     2581 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     2641 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     2701 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     2761 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     2821 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     2881 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     2941 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     3001 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     3061 cgcgcacccg ggttctagag gaagcacctc cggcgataac gattacaagg atgatgatga
     3121 taaggccagc ggcaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     3181 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     3241 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3301 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3361 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3421 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     3481 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3721 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3781 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3841 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3901 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3961 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4021 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4081 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4141 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4201 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4261 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4321 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4381 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4441 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4501 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4561 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4621 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4681 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4741 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4801 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4861 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4921 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4981 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5041 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5101 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5161 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5221 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5281 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5341 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5401 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5461 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5521 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5581 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5641 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5701 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5761 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5821 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5881 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5941 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6001 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6061 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
