LOCUS       sequence-238687-        6602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G46350 N7_pECIA14 should be used with bait vector 
            AT3G46350 N7_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6602)
  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 6602)
  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: AT3G46350 N7_pECIA14
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2343..2366
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4041..4064
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4080..4097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4120..4137)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4170..4304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4194..4213)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4248..4267
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4448..4464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4448..4464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4461..4483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4472..4488
                     /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(4496..4502,4503..4520,4521..4526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4541..4562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4679..4696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4850..5438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4930..4949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5609..6400,6401..6469))
                     /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     6232..6251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6470..6574)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggcaaga aggattcata agtttggatt gtgggttagc tccgactgag ccgtcccctt
     1021 ataccgagcc ggtaactacg ttacagtact catcggattc aaacttcatc caaagtggga
     1081 aacttggtag aattgataca agtctacaga catttttcct aaagcaacaa acaacgttaa
     1141 gatactttcc agacggaata cgcaattgtt acaatcttac ggtcaagcag ggaaccaact
     1201 atttgatcag agctaggttc acatatggga attatgatgg tcgtaatatg tctcctacct
     1261 ttgatctata cttgggcccc aatttgtgga aaagaataga catgacaaag ttacaaaaca
     1321 aagttagtac attggaggag attacataca ttccattatc aaactctttg gatgtgtgtc
     1381 ttgttaagac aaacacgacc attccattta tatcggcctt agaattaagg cctttaccaa
     1441 gtaacagtta tatcactacg gccggttcat tgaggacgtt cgtgcggttt tgtttcagca
     1501 attcagtgga agatatacgg ttcccgatgg atgtccatga tagaatgtgg gaatcatact
     1561 ttgatgacga ctggacgcag atttccacta gtcttacagt gaacacatcc gatagttttc
     1621 gtctgccaca ggctgccctc ataacggctg caacacctgc gaaggatggt ccatcatata
     1681 taggtatcac gttttctacc tcctctgaag aaaggttttt catatacctc catttctccg
     1741 aagttcaagc tttacgtgcc aacgagacta gagaattcaa catttcgatt aacggagaat
     1801 ctgttgccga cttgtataga cctttgtcaa gaactcaaag ctctacgcat ccacctatga
     1861 ttaacgccat tgaaattttc cttgtttccg aacttctaca gtctgaaaca tacgagaatg
     1921 atgtgattgc tataaaaaaa atcaaagaca catatggatt gcagctaatc tcatggcaag
     1981 gtgacccatg tgttcctcga ctttataagt gggacggtct tgattgtaca gacacagaca
     2041 catatatagc accaaggatc acttccttaa aattatcttc aaaggggtta acgggaacca
     2101 tagcagctga tattcaatat ctcacctctc tagaaaagtt ggacttgtca gataacaagt
     2161 tggtgggagt agtgcctgaa tttctagcta acatgaaatc actaatgttc ataaacttga
     2221 ccaagaatga tctgcatggt tcaattccac aagcacttcg cgatagagaa aagaaaggac
     2281 tgaaaatctt gtttgatggt gataagaatg atccatgttt atctacctct tgtaacccaa
     2341 agctggaggt gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga
     2401 tgctgcgcga gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc
     2461 accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca
     2521 tgcagccctc tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct
     2581 ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag
     2641 ccgccaagaa cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg
     2701 ccaggatcct aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg
     2761 accgcttccc atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca
     2821 gtggagccac agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct
     2881 tgagtgcagc cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg
     2941 tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc
     3001 agcacgcctc gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg
     3061 ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct
     3121 ccaacatgga cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa
     3181 ccccagaccc tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga
     3241 atctggtgca ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg
     3301 agctcatgca ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg
     3361 gagacatgaa atacgagatc caccgagact ccacactgga cccctccctg atggagatga
     3421 cagaggctgc cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg
     3481 gtggtcgcat cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga
     3541 tcatgttcga cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga
     3601 gcctcgtcac tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga
     3661 gctccatctt cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc
     3721 tatacggaaa cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga
     3781 gcgagagcgg gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc
     3841 acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg
     3901 tgcaggagca gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca
     3961 ccgcctgcga cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag
     4021 gaagcacctc cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc
     4081 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg
     4141 cctgagctcg ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt
     4201 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4261 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4321 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     4381 tctacaaatg tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt
     4441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6601 ac
//
