LOCUS       sequence-270157-        6497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G46370 N8_pECIA14 should be used with bait vector 
            AT3G46370 N8_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  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 6497)
  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: AT3G46370 N8_pECIA14
FEATURES             Location/Qualifiers
     source          1..6497
                     /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             1793..1816
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3491..3514
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3530..3547
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3570..3587)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3620..3754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3644..3663)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3698..3717
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3898..3914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3898..3914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3911..3933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3922..3938
                     /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(3946..3952,3953..3970,3971..3976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3991..4012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4129..4146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4300..4888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4380..4399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5059..5850,5851..5919))
                     /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     5682..5701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5920..6024)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6092..6110
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6148..6170)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6270..6289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6483..6497,1..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 atgcgaaact gttacaatct
      541 tagtgtgcat aaagaaacaa agtatttgat tagggttaca agtaactatg gaaattacga
      601 tggtcgtaac gaacctccta gatttgattt gtacttaggc cctaacttct gggtaacaat
      661 agatttgggg aagcatgtaa atggtgatac atggaaggag attattcata tcccaaagtc
      721 aaactctcta gatgtgtgtc ttatcaaaac aggtacaaca acgccaatta tatcgacctt
      781 ggaactaagg tctctaccaa agtatagtta caacgcaata tcaggttctt tgaagtcgac
      841 actgagggca ttcctaagcg aatcaacaga agttataagg tacccaaatg atttctacga
      901 tcggatgtgg gttccacatt ttgagacgga atggaagcaa atttctacca atctcaaagt
      961 gaacagctcc aatggttatc ttctgccaca ggatgtgctc atgaccgccg caatacctgt
     1021 gaatactagt gcacgattga gttttaccga gaatcttgag ttccctcatg atgaacttta
     1081 cctgtacttc cacttctctg aggtccaagt cttacaggcc aaccagagta gagagttcag
     1141 cattttgtgg aacggaatgg ttatttaccc agattttatc cctgactatc ttggtgccgc
     1201 cactgtgtac aacccgtcac catctctttg tgaagtaggg aaatgcttat tagagcttga
     1261 aagaactcag aaatcaactc tcccgcctct ccttaacgcc attgaagttt tcacagtgat
     1321 gaattttcca cagtctgaga caaacgatga tgatgtaatt gctataacaa agatcaaaga
     1381 cacacacaga ttgaatagaa cctcctggca aggagatcca tgcgtccctc aactgttttc
     1441 gtgggccggt ctaagttgca ttgacacgaa tgtatctacc ccaccaagaa tcatttccct
     1501 aaacttgtct tcaagtgggt taacaggaaa catagcaact ggaattcaaa atctgaccaa
     1561 actgcaaaag ctggacttgt caaataacaa tttgacagga gtagtgcccg aatttctagc
     1621 taacatgaaa tcgttgctgt tcatagactt gaggaagaac aaactgaatg gttcaatacc
     1681 aaagactcta cttgatagaa aaaagaaagg actacaatta tttgttgatg gcgatgatga
     1741 taagggcgat gataataagt gtttatccgg ttcatgtgtc ccaaaaatga aactggaggt
     1801 gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     1861 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     1921 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca tgcagccctc
     1981 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2041 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2101 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2161 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     2221 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2281 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2341 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2401 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     2461 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     2521 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     2581 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2641 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2701 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2761 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     2821 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     2881 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     2941 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3001 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3061 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3121 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3181 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3241 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3301 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3361 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3421 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag gaagcacctc
     3481 cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc atcatcacca
     3541 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     3601 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     3661 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3721 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3781 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3841 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     3901 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3961 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4021 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5101 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5161 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5221 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5281 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5341 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5401 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5461 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5521 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5581 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5641 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5701 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5761 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5821 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5881 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5941 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6001 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6061 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6121 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6181 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6241 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6301 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6361 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6421 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6481 ttcccagtca cgacgtt
//
