LOCUS       sequence-270262-        5840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G64210 I9_pECIA14 should be used with bait vector 
            AT1G64210 I9_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5840)
  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 5840)
  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: AT1G64210 I9_pECIA14
FEATURES             Location/Qualifiers
     source          1..5840
                     /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             1136..1159
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2834..2857
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2873..2890
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2913..2930)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2963..3097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2987..3006)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3041..3060
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3241..3257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3241..3257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3254..3276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3265..3281
                     /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(3289..3295,3296..3313,3314..3319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3334..3355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3472..3489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3643..4231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3723..3742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4402..5193,5194..5262))
                     /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     5025..5044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5263..5367)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5435..5453
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5491..5513)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5613..5632
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5826..5840,1..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 tctcagactc ttgaagacga
      541 caagaaagct ttgcttcatt tcttgtctag tttcaattct tctcgtctcc attggaacca
      601 gagctcagac gtttgtcata gctggaccgg tgttacctgc aacgaaaacg gagatcgaat
      661 tgtatccgtt agattgcctg cggttggatt caacggtttg attcctcctt tcaccatcag
      721 tcgtctctct tctctcaagt ttctaagcct tagaaagaac catttcaccg gagacttccc
      781 ttctgatttc acaaatctca agagcttgac tcatctctac ctacaacata accacctctc
      841 aggtccacta ctagctatct tctcggagtt gaagaatctt aaggttcttg atctgtctaa
      901 caatggattc aacggaagca tcccaacctc tctttcagga ttgactagcc ttcaagtctt
      961 gaatcttgca aacaattcat tctcagggga aattccaaat cttcatcttc caaaactgag
     1021 tcagatcaac ttgtccaaca acaagctcat cggtacaata ccaaagtctc ttcaaagatt
     1081 ccaaagttca gctttttctg gcaacaacct taccgagagg aagaaacaac gcaagctgga
     1141 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     1201 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     1261 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     1321 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     1381 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     1441 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     1501 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     1561 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     1621 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     1681 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     1741 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     1801 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     1861 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     1921 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     1981 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2041 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     2101 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2161 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2221 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2281 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2341 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     2401 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     2461 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     2521 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     2581 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     2641 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     2701 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     2761 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     2821 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     2881 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     2941 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3001 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3061 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3121 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3181 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3301 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3421 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3961 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4081 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4141 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4201 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4261 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4321 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4381 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4441 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4501 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4561 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4621 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4681 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4741 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4801 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4861 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4921 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4981 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5041 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5101 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5161 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5221 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5281 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5341 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5401 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5461 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5521 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5581 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5641 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5701 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5761 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5821 gttttcccag tcacgacgtt
//
