LOCUS       sequence-274544-        6440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT3G53590 X063_pECIA14 should be used with bait vector 
            AT3G53590 X063_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  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 6440)
  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: AT3G53590 X063_pECIA14
FEATURES             Location/Qualifiers
     source          1..6440
                     /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             1736..1759
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3434..3457
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3473..3490
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3513..3530)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3563..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3641..3660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3841..3857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3841..3857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3854..3876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3865..3881
                     /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(3889..3895,3896..3913,3914..3919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3934..3955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4072..4089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4243..4831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4323..4342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5002..5793,5794..5862))
                     /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     5625..5644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5863..5967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6035..6053
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6091..6113)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6213..6232
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6426..6440,1..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 tccttgaaac tcctactgtt
      541 gaatgggaac aagttcacgg gtagtttacc ccctgaactt ggtaaccttc aaaacctaaa
      601 caggcttcaa gtagatgaga ataacataac aggttcagtt ccattttcat ttggaaactt
      661 gagaagcata aaacatctcc acttaaacaa taacacgata agtggagaaa ttcctgttga
      721 gctatccaaa ttgcccaagc ttgttcacat gatactggac aacaataact tgactgggac
      781 attgccgcta gagttagctc aattgccatc cttgaccata cttcagctgg ataacaacaa
      841 cttcgaaggt tcaactattc cagaggctta cggacacttt tctagattag taaaattaag
      901 tttaaggaac tgtggactgc aaggatctat tcctgattta agcaggatag aaaatcttag
      961 ctatctagac ctaagttgga accacctcac aggcactata ccagaatcca agctttctga
     1021 caatatgaca acaattgaac tatcttataa ccacctaact ggatctatac cacaaagttt
     1081 ctcagatttg aactctcttc aattactgtc acttgagaac aacagtttat ctggttctgt
     1141 cccaacagaa atttggcagg acaagtcttt cgagaataac aagctgcaag ttgatctcag
     1201 gaacaacaac ttctctgatg ccacaggaaa cctgagaaca ccagacaatg ttaccctata
     1261 tcttcgtggc aacccaatat gcaagagtac aagcattccc atggttacac aattcttcga
     1321 atacatctgc ggcgaaaaaa agcagacatc tacaaactca aacacaccct gcagtaatgt
     1381 gtcatgtccc tttgagaatg tgaaagtatc cccaggaatc tgcttgtgca cagcacctct
     1441 ttcgattgat tataggctaa agagccctag cttctttttc tttactccat atattgaacg
     1501 ccaattcagg gagtatatta cctcgtccct ccaactggag actcatcaac tagcgattga
     1561 tagacttgtc gatgagaaca ggctgcgtcc aagaatgtac ttaaagctag tccccaaagg
     1621 cagaataaca tttaacaaga gtgaagtcat acgtatacgg gacaggttta tgtcatggag
     1681 ttttaataaa actgactttt ttggaccata tgagctgctg gacttccctc ttcaactgga
     1741 ggtgctgttc cagggaccaa gcggcaatcc cggcggcgat ctggcccccc agatgctgcg
     1801 cgagctgcag gagaccaatg ccgccctgca ggacgtgcgc gagctgctgc gccaccgcgt
     1861 gaaggagatc accttcctga agaacaccgt gatggagtgc gacgcctgcg gcatgcagcc
     1921 ctctagaggc tccggctcca tcatcccagt tgaggaggag aacccggact tctggaaccg
     1981 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     2041 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     2101 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     2161 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     2221 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     2281 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     2341 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     2401 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     2461 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     2521 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     2581 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2641 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     2701 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2761 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2821 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2881 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2941 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     3001 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     3061 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     3121 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     3181 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     3241 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     3301 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     3361 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggttcta gaggaagcac
     3421 ctccggcgat aacgattaca aggatgatga tgataaggcc agcggcaccg gtcatcatca
     3481 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     3541 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     3601 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3661 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3721 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3781 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3841 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3901 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3961 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4021 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4081 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4141 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4201 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4261 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4321 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4381 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4441 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4501 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4561 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4621 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4681 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4741 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4801 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4861 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4921 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6001 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6061 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6241 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6421 gttttcccag tcacgacgtt
//
