LOCUS       sequence-274066-        6695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G62950 X004_pECIA14 should be used with bait vector 
            AT1G62950 X004_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  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 6695)
  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: AT1G62950 X004_pECIA14
FEATURES             Location/Qualifiers
     source          1..6695
                     /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             1991..2014
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3689..3712
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3728..3745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3768..3785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3818..3952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3842..3861)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3896..3915
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4096..4112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4096..4112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4109..4131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4120..4136
                     /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(4144..4150,4151..4168,4169..4174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4189..4210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4327..4344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4498..5086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4578..4597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5257..6048,6049..6117))
                     /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     5880..5899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6118..6222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6290..6308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6346..6368)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6468..6487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6681..6695,1..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 tcggattcta ttattaccga
      541 acgagagatt ctacttcaat tcaaagacaa catcaacgat gatccgtaca acagtttagc
      601 ttcttgggtc tctaatgctg acctctgtaa cagcttcaat ggtgtatctt gtaaccaaga
      661 aggattcgtc gaaaagatcg ttctttggaa cactagtctc gccggaaccc taacgccggc
      721 attgtctggt ttaacttctt taagggtttt gactctgttt ggcaacagga ttacaggtaa
      781 tttacctttg gattacttaa agttacagac tttatggaag atcaatgtta gttccaatgc
      841 attgtctggt ttggttcctg aatttatcgg tgatttgcct aatttaaggt tcttggattt
      901 gtcgaaaaat gcgttctttg gagagattcc aaattctctg tttaaattct gttacaagac
      961 aaagtttgtt tcattgtctc ataacaatct ctcaggatca atccctgaat caatcgtaaa
     1021 ctgtaacaat ctcattggtt tcgatttctc ttacaatggc attaccggtt tgttgcctcg
     1081 gatctgcgat attccggttc ttgagtttgt atctgttaga agaaatttgt tgtctggtga
     1141 tgtgtttgag gagatatcga agtgtaagag attgagtcat gtcgatatag gaagcaattc
     1201 tttcgatgga gtagcgtctt tcgaggttat tggattcaag aacctgactt attttaatgt
     1261 gtctggaaac cggtttagag gggagattgg tgagattgtt gattgtagtg aaagtttgga
     1321 gtttttggat gcttcttcga atgagttaac tggtaatgta cctagcggta taaccggatg
     1381 caaaagtctt aagcttttgg acttggaatc gaataggctc aatgggagtg tacctgtagg
     1441 tatggggaag atggagaagc tctctgtgat caggttaggc gataacttta tagatgggaa
     1501 gctaccactt gagctcggaa atcttgagta tctacaggtt ttgaatttgc ataatctcaa
     1561 tctcgttggt gaaataccag aggatttatc aaactgcaga ctacttcttg aactagatgt
     1621 ttctggaaat ggtttggaag gagagatccc taagaatcta ttgaacttaa caaacttaga
     1681 gattcttgat ctgcatcgga accgaatcag tggaaacatt ccgcctaatc tcgggagcct
     1741 ctcgagaatc cagttcctcg atctttcaga gaatttgctt tctggtccta tcccgtcttc
     1801 cctcgagaat ttgaaaagac tgactcattt caatgtttct tacaacaacc tctccggcat
     1861 tatcccaaag attcaagctt cgggagcttc ttcgttttcc aacaaccctt tcctctgcgg
     1921 tgatcctctt gaaacaccgt gcaatgctct cagaaccgga tcaagatcaa gaaaaaccaa
     1981 agctctaagc ctggaggtgc tgttccaggg accaagcggc aatcccggcg gcgatctggc
     2041 cccccagatg ctgcgcgagc tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct
     2101 gctgcgccac cgcgtgaagg agatcacctt cctgaagaac accgtgatgg agtgcgacgc
     2161 ctgcggcatg cagccctcta gaggctccgg ctccatcatc ccagttgagg aggagaaccc
     2221 ggacttctgg aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc
     2281 acagacagcc gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt
     2341 gacagctgcc aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct
     2401 ggccatggac cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt
     2461 gccagacagt ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac
     2521 cattggcttg agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt
     2581 catctccgtg atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac
     2641 acgagtgcag cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta
     2701 ctcggacgcc gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca
     2761 gctcatctcc aacatggaca ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg
     2821 catgggaacc ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga
     2881 cgggaagaat ctggtgcagg aatggctggc gaagcgccag ggtgcccggt atgtgtggaa
     2941 ccgcactgag ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt
     3001 tgagcctgga gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat
     3061 ggagatgaca gaggctgccc tgcgcctgct gagcaggaac ccccgcggct tcttcctctt
     3121 cgtggagggt ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac
     3181 tgagacgatc atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga
     3241 cacgctgagc ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct
     3301 gcgagggagc tccatcttcg ggctggcccc tggcaaggcc cgggacagga aggcctacac
     3361 ggtcctccta tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt
     3421 taccgagagc gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga
     3481 agagacccac gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt
     3541 tcacggcgtg caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga
     3601 gccctacacc gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg
     3661 ttctagagga agcacctccg gcgataacga ttacaaggat gatgatgata aggccagcgg
     3721 caccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     3781 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     3841 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3901 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3961 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4021 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     4081 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4141 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4201 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4261 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4321 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4381 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4441 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4501 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4561 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4621 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4681 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4741 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4801 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4861 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4921 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4981 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5041 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5101 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5161 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5221 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5281 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5341 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5401 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5461 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5521 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5581 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5641 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5701 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5761 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5821 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5881 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5941 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6001 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6061 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6121 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6181 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6241 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6301 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6361 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6421 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6481 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6541 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6601 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6661 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
