LOCUS       sequence-297021-        6455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G75640 X061_pECIA2 should be used with prey vector 
            AT1G75640 X061_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6455)
  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 6455)
  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: AT1G75640 X061_pECIA2
FEATURES             Location/Qualifiers
     source          1..6455
                     /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             2708..2731
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3437..3478
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3488..3505
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3528..3545)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3578..3712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3602..3621)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3656..3675
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3856..3872)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3856..3872)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3869..3891)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3880..3896
                     /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(3904..3910,3911..3928,3929..3934))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3949..3970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4087..4104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4258..4846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4338..4357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5017..5808,5809..5877))
                     /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     5640..5659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5878..5982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6050..6068
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6106..6128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6228..6247
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6441..6455,1..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 tcatcagaga ctcaagcact
      541 cacatctttc aaactctctc tccacgaccc tctaggagct ctcgagtcct ggaatcaatc
      601 atctccatca gctccatgcg actggcacgg cgtttcttgc ttctccggcc gtgtccgtga
      661 gcttcgtctt cctcgtctcc acctcaccgg ccacctcagt cctcgactcg gtgagttgac
      721 tcagctccgt aaactaagtc ttcacaccaa cgacataaac ggcgccgttc cttcgtctct
      781 ctctcgttgt gtcttcctta gagctcttta tcttcactac aactccttct ccggcgattt
      841 tccaccggag atactgaatc tacgaaacct ccaggtcctc aacgctgctc acaactcact
      901 caccggaaat ctatccgacg tcacagtctc gaagagcctt cgttacgtcg atttgtcttc
      961 caatgcaata tccggtaaga ttccggcgaa tttctcagcg gattcatctc tccagctcat
     1021 caacctctcg ttcaaccact tctccggtga gatcccggct acattgggac aactacaaga
     1081 cctcgagtat ctctggctag actcaaacca attacaaggc acaataccat ccgcattagc
     1141 caactgttca tcgcttatac acttcagcgt aaccggtaat cacctcaccg gtttaattcc
     1201 ggttacttta ggcacgattc gaagcctgca ggtgatttca ttatcggaga attcatttac
     1261 cggtacagtt ccggttagtc ttctatgtgg ttattcaggt tacaattcct ccatgcgaat
     1321 cattcaatta ggagttaata acttcacggg aatagctaag ccgtcaaatg cagcttgcgt
     1381 gaaccctaat ttggagattc tcgacattca cgagaatcgc attaatggcg attttcccgc
     1441 gtggttgact gatttgacct ctttagtggt tttggatatt tctggaaacg gtttctccgg
     1501 tggtgttacg gctaaagttg ggaacttgat ggcgcttcag gagctgagag tagctaacaa
     1561 ttcacttgtt ggagaaattc cgacgagtat ccgtaattgc aaatcgttaa gagtcgttga
     1621 tttcgaagga aacaagttct cgggtcagat tccgggtttt ctaagtcagt tacggagcct
     1681 gacaacgata tctcttggaa gaaacgggtt ctcgggtcgg attccatcgg atcttctcag
     1741 cttgtacgga ctcgagacat tgaatctcaa tgaaaatcat ctcaccggag caataccgtc
     1801 ggagataaca aagctagcta atcttaccat ccttaatctc agtttcaaca gattctccgg
     1861 cgaagttccg tccaatgtgg gtgatctgaa aagcttgagt gttctcaaca taagtggatg
     1921 cgggttaacg ggtcggatac cggttagcat tagcggatta atgaagcttc aggttttgga
     1981 tataagtaaa cagagaatct ctggtcagtt accagtcgaa ctatttggtt taccggatct
     2041 tcaagtggtt gctcttggaa acaacttgct cggcggcgtc gtaccggaag gtttcagcag
     2101 cttggttagt cttaagtact taaatctcag ttccaactta ttctccggac atatcccaaa
     2161 gaactatgga tttctcaagt cattacaagt gctttcgttg tctcacaatc gaatatccgg
     2221 tacaatcccg ccggagattg ggaattgttc cagtcttgag gttcttgagc ttggttctaa
     2281 tagtttaaag ggtcacattc cggtttacgt ttccaagttg tctcttttga agaagcttga
     2341 tttgagtcac aacagtttaa cagggagtat tccggatcag atttctaagg attcttctct
     2401 agaatctctc ctgcttaatt cgaattctct atcgggtcga ataccggaat ctttatcaag
     2461 attgacgaat ctaaccgcgt tggatctttc gtcgaacaga ttgaattcca ctataccatc
     2521 aagtctctct cggcttcgtt tcttgaacta tttcaactta tcaagaaaca gtttggaagg
     2581 ggagattcca gaagctctag ctgcgcgttt cactaacccg accgtctttg tcaagaaccc
     2641 gggactctgc ggtaaaccgc ttggtataga atgtccaaac gttcgaagga gaagaagaag
     2701 aaagctactg gaggtgctgt tccagggacc agggagcgac aaaactcaca catgcccacc
     2761 gtgcccagca cctgaactcc tggggggacc gtcagtcttc ctcttccccc caaaacccaa
     2821 ggacaccctc atgatctccc ggacccctga ggtcacatgc gtggtggtgg acgtgagcca
     2881 cgaagaccct gaggtcaagt tcaactggta cgtggacggc gtggaggtgc ataatgccaa
     2941 gacaaagccg cgggaggagc agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt
     3001 cctgcaccag gactggctga atggcaagga gtacaagtgc aaggtctcca acaaagccct
     3061 cccagccccc atcgagaaaa ccatctccaa agccaaaggg cagccccgag aaccacaggt
     3121 gtacaccctg cccccatccc gggaggagat gaccaagaac caggtcagcc tgacctgcct
     3181 ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga
     3241 gaacaactac aagaccacgc ctcccgtgct ggactccgac ggctccttct tcctctacag
     3301 caagctcacc gtggacaaga gcaggtggca gcaggggaac gtcttctcat gctccgtgat
     3361 gcatgagggt ctgcacaacc actacacgca gaagagcctc tccctgtctc cgggtaaatc
     3421 tagagggccc ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     3481 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     3541 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     3601 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3661 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3721 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     3781 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca ggcatgcaag
     3841 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3901 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3961 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4021 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4081 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4141 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4201 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4261 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4321 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4381 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4441 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4501 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4561 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4621 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4681 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4741 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4801 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4861 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4921 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4981 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5041 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5101 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5161 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5221 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5281 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5341 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5401 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5461 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5521 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5581 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5641 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5701 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5761 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5821 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5881 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5941 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6001 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6061 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6121 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6181 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6241 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6301 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6361 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6421 ttgggtaacg ccagggtttt cccagtcacg acgtt
//
