LOCUS       sequence-270425-        6947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G10020 X022_pECIA14 should be used with bait vector 
            AT5G10020 X022_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6947)
  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 6947)
  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: AT5G10020 X022_pECIA14
FEATURES             Location/Qualifiers
     source          1..6947
                     /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             2243..2266
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3941..3964
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3980..3997
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4020..4037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4070..4204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4094..4113)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4148..4167
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4348..4364)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4348..4364)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4361..4383)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4372..4388
                     /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(4396..4402,4403..4420,4421..4426))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4441..4462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4579..4596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4750..5338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4830..4849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5509..6300,6301..6369))
                     /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     6132..6151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6370..6474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6542..6560
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6598..6620)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6720..6739
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6933..6947,1..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 accgagacgg agctccgatc
      541 tctactcgag ttccgtaaag ggatccgtga cgagacctca caccaacgaa tctcatggtc
      601 agatacaagc agcctcactg atccatctac ttgccctaat gactggcctg gtatctcttg
      661 cgaccctgaa accggttcaa tcatcgccat taatctcgac cgtcgtggtc tctccggcga
      721 actcaaattc agcactctct ctggtcttac tcgtcttcgt aacctctctt tatccggaaa
      781 cagcttctcg ggtcgggtcg ttccttcatt gggtggaatc tcgtcgcttc aacacttaga
      841 tctgtcggat aacggtttct acggacctat accgggtcgg atctcggagc tttggagttt
      901 gaatcatctc aatttgtctt ccaacaagtt cgaaggtggg tttcctagtg gctttcggaa
      961 tcttcagcag ctaagatctt tggatttgca taagaacgag atttggggag atgttggtga
     1021 gattttcact gagcttaaga atgttgagtt cgttgatttg agctgtaacc gtttcaacgg
     1081 tggcttatct ttaccaatgg agaacatttc cagcatttcc aacactctcc gtcacttgaa
     1141 tttgagccac aatgcgttaa atggcaagtt tttcagtgag gagtccattg gttcgttcaa
     1201 gaatctcgag attgttgact tagagaataa ccagatcaat ggtgagttac cgcattttgg
     1261 atcacagccg agtttgagaa tattgaagct tgctcgtaac gagttgttcg gtttagtacc
     1321 tcaagagttg ttgcagagtt cgattccttt gctagagcta gatctaagtc gaaacggttt
     1381 tacaggatca attagtgaaa tcaattcatc aacgttaact atgttgaatc tttcttctaa
     1441 cggcctgtct ggggatcttc cgtcgagttt taaaagttgc tcagtgatag atttgagtgg
     1501 gaatactttc tctggcgacg tgtctgttgt tcaaaagtgg gaagctactc cagatgttct
     1561 tgatcttagc tcaaacaact tgtcgggaag cttgccgaac tttacttctg cattttcaag
     1621 attaagtgtg ttgagcatta ggaataattc tgtctctggt agcttgcctt ctctgtgggg
     1681 tgattctcaa ttctctgtga ttgatctcag ttcaaataag ttcagtggct tcattccagt
     1741 gagtttcttc acttttgcaa gcttgagaag tttgaatctt tctaggaata acttggaggg
     1801 gccaattcct tttcgaggtt cacgtgcaag cgaacttttg gttctaaatt cttatccgca
     1861 gatggagtta cttgaccttt caaccaattc tttgacgggt atgctgcctg gggatatagg
     1921 tacaatggaa aaaattaaag tgctgaatct tgccaataat aagctgtccg gggaacttcc
     1981 aagcgacctc aacaagctta gtggcctttt gttccttgac ttgtcaaaca atactttcaa
     2041 gggtcaaatt cccaataagc ttccttccca aatggtggga ttcaatgtgt cgtataacga
     2101 cctatcaggc ataattccgg aagaccttag aagctacccg ccttcttcat tttatcctgg
     2161 taactccaag ctaagtctcc caggacggat tccagcggat tcttctggtg atctgtcttt
     2221 acctgggaaa aaacatcatt cgctggaggt gctgttccag ggaccaagcg gcaatcccgg
     2281 cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg ccctgcagga
     2341 cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat
     2401 ggagtgcgac gcctgcggca tgcagccctc tagaggctcc ggctccatca tcccagttga
     2461 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     2521 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     2581 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     2641 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     2701 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     2761 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     2821 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     2881 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     2941 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     3001 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     3061 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     3121 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     3181 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     3241 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     3301 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     3361 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     3421 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     3481 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     3541 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     3601 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     3661 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     3721 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     3781 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     3841 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     3901 cgcgcacccg ggttctagag gaagcacctc cggcgataac gattacaagg atgatgatga
     3961 taaggccagc ggcaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     4021 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     4081 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4141 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4201 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4261 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     4321 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4381 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4441 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4501 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4561 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4621 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4681 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4741 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4801 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4861 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4921 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4981 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5041 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5101 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5161 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5221 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5281 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5341 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5401 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5461 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5521 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5581 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5641 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5701 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5761 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5821 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5881 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5941 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6001 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6061 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6121 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6181 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6241 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6301 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6361 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6421 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6481 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6541 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6601 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6661 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6721 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6781 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6841 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6901 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
