LOCUS       sequence-238695-        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G59650 O4_pECIA14 should be used with bait vector 
            AT5G59650 O4_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  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 6659)
  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: AT5G59650 O4_pECIA14
FEATURES             Location/Qualifiers
     source          1..6659
                     /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             2400..2423
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4098..4121
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4137..4154
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4177..4194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4227..4361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4251..4270)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4305..4324
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4505..4521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4505..4521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4518..4540)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4529..4545
                     /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(4553..4559,4560..4577,4578..4583))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4598..4619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4736..4753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4907..5495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4987..5006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5666..6457,6458..6526))
                     /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     6289..6308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6527..6631)
                     /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 tcgggagttt ggattgcggg ttacccatga ctgaaccgtc ttcttatact gagtcagtaa
     1021 ccggactacg gttctcgtct gatgctgaat tcatccagac tggtgaaagt ggtaaaatcc
     1081 aggcgagtat ggagaatgac taccttaagc cgtacacgag gctgaggtat tttccagagg
     1141 agagaaggaa ctgctatagt ttgagtgttg acaagaacag gaaatatttg atcagggcta
     1201 ggttcattta tgggaattac gatggtcgta actctaaccc gatatttgaa ctgcatctag
     1261 gacctaatct gtgggcaact atagatttgc aaaagtttgt gaatggtaca atggaggaga
     1321 tccttcacac tccaacatca aactctttga acgtttgtct tgttaagaca gggacaacta
     1381 cacccttgat ctcagccttg gaattacggc cattaggaaa taattcttat ctcacagatg
     1441 gttctttgaa tcttttcgtt cggatatatc tcaacaagac agatggcttt cttcggtacc
     1501 cggatgatat ttatgatcgc agatggcata attactttat ggtggatgat tggacccaaa
     1561 ttttcaccac tctcgaagtg accaatgata acaattatga accaccaaaa aaggctcttg
     1621 cagcggctgc cacgccttct aatgccagcg cgccattgac aatcagttgg ccaccagaca
     1681 atcctggtga ccaatattac ttgtacagcc acttctctga gatacaagat ttacagacca
     1741 atgataccag agaatttgac atattatggg atggagctgt tgttgaagaa ggtttcatcc
     1801 ctccaaagtt aggggtgact actatccata atctttctcc agtgacatgc aaaggagaaa
     1861 attgcattta ccagctaata aaaacctcga ggtcaactct tccttctcta cttaacgctc
     1921 ttgaaatcta cacagttatc caatttccac ggtcggaaac aaatgaaaat gatgtggttg
     1981 ctgttaaaaa tatcgaagct gcctataaat tgagtagaat caggtggcaa ggagatcctt
     2041 gcgtccctca aaagtatgcg tgggatggtt tgaactgtag caacaacaca gatgtttcta
     2101 aacctccaag agtattatcc ttaaatttgt cttcgagcgg tttaactgga atcattgcgg
     2161 ccgcaattca aaaccttaca catttagaaa aactggactt gtcaaataac acactaactg
     2221 gagtggtgcc ggaatttcta gcccagatga aatcgttggt gatcataaac ttaagcggca
     2281 acaatcttag tggtcccctt cctcaaggtc ttcgaagaga gggactagaa ttactagttc
     2341 aaggcaaccc aaggctttgt ctctctggtt cctgcacaga aaaaaactcg aagaagaaac
     2401 tggaggtgct gttccaggga ccaagcggca atcccggcgg cgatctggcc ccccagatgc
     2461 tgcgcgagct gcaggagacc aatgccgccc tgcaggacgt gcgcgagctg ctgcgccacc
     2521 gcgtgaagga gatcaccttc ctgaagaaca ccgtgatgga gtgcgacgcc tgcggcatgc
     2581 agccctctag aggctccggc tccatcatcc cagttgagga ggagaacccg gacttctgga
     2641 accgcgaggc agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg
     2701 ccaagaacct catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca
     2761 ggatcctaaa agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc
     2821 gcttcccata tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg
     2881 gagccacagc cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga
     2941 gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga
     3001 tgaatcgggc caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc
     3061 acgcctcgcc agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg
     3121 acgtgcctgc ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca
     3181 acatggacat tgacgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc
     3241 cagaccctga gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc
     3301 tggtgcagga atggctggcg aagcgccagg gtgcccggta tgtgtggaac cgcactgagc
     3361 tcatgcaggc ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag
     3421 acatgaaata cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag
     3481 aggctgccct gcgcctgctg agcaggaacc cccgcggctt cttcctcttc gtggagggtg
     3541 gtcgcatcga ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca
     3601 tgttcgacga cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc
     3661 tcgtcactgc cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct
     3721 ccatcttcgg gctggcccct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat
     3781 acggaaacgg tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg
     3841 agagcgggag ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg
     3901 caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc
     3961 aggagcagac cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg
     4021 cctgcgacct ggcgcccccc gccggcacca ccgacgccgc gcacccgggt tctagaggaa
     4081 gcacctccgg cgataacgat tacaaggatg atgatgataa ggccagcggc accggtcatc
     4141 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct
     4201 gagctcgctg atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg
     4261 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     4321 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     4381 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     4441 acaaatgtgg tatggctgat tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat
     4501 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4561 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4621 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4681 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4741 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4801 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5101 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5341 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
