LOCUS       sequence-297042-        5483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G53590 X063_pECIA2 should be used with prey vector 
            AT3G53590 X063_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5483)
  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 5483)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..5483
                     /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             2465..2506
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2516..2533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2556..2573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2606..2740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2630..2649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2684..2703
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2884..2900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2884..2900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2897..2919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2908..2924
                     /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(2932..2938,2939..2956,2957..2962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2977..2998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3115..3132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3286..3874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3366..3385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4045..4836,4837..4905))
                     /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     4668..4687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4906..5010)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5078..5096
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5134..5156)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5256..5275
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5469..5483,1..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 cagggaccag ggagcgacaa aactcacaca tgcccaccgt gcccagcacc
     1801 tgaactcctg gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat
     1861 gatctcccgg acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga
     1921 ggtcaagttc aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg
     1981 ggaggagcag tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga
     2041 ctggctgaat ggcaaggagt acaagtgcaa ggtctccaac aaagccctcc cagcccccat
     2101 cgagaaaacc atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc
     2161 cccatcccgg gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt
     2221 ctatcccagc gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa
     2281 gaccacgcct cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt
     2341 ggacaagagc aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgagggtct
     2401 gcacaaccac tacacgcaga agagcctctc cctgtctccg ggtaaatcta gagggccctt
     2461 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     2521 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     2581 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     2641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2761 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2821 caaatgtggt atggctgatt atgatcagtc gacctgcagg catgcaagct tggcgtaatc
     2881 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2941 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3001 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3061 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3721 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3961 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4201 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4261 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4321 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4381 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4441 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4501 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4561 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4621 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4681 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4741 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4801 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4861 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4921 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4981 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5041 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     5101 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5161 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5221 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     5281 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     5341 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5401 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     5461 agggttttcc cagtcacgac gtt
//
