LOCUS       sequence-296504-        5687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G51890 M5_pECIA2 should be used with prey vector 
            AT1G51890 M5_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5687)
  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 5687)
  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: AT1G51890 M5_pECIA2
FEATURES             Location/Qualifiers
     source          1..5687
                     /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             1940..1963
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2669..2710
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2720..2737
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2760..2777)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2810..2944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2834..2853)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2888..2907
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3088..3104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3088..3104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3101..3123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3112..3128
                     /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(3136..3142,3143..3160,3161..3166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3181..3202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3319..3336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3490..4078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3570..3589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4249..5040,5041..5109))
                     /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     4872..4891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5110..5214)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5282..5300
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5338..5360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5460..5479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5673..5687,1..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 tcaggattca taagcttaga
      541 ttgtggtttg gtgcctacgg aaattactta tgtggaaaag tcgacgaata taacatacag
      601 atcagacgca acttacatcg acagtggagt tcccgggaag atcaatgaag tgtacagaac
      661 acagtttcag caacaaattt gggccttgag aagcttccct gagggtcaaa gaaattgtta
      721 caacttcagt ctcaccgcaa aacgtaagta tctaatcaga ggaaccttta tctatgggaa
      781 ttatgacggt ttgaatcaac tcccgagctt tgatctttac atcggtccaa acaaatggac
      841 ctctgtttcg atccccggag tgagaaatgg ttcagtctcc gagatgatcc atgtcttaag
      901 acaagaccat cttcaaattt gtcttgtgaa aacaggagaa actacaccgt ttatttcttc
      961 attggaactt cgtcctttga acaataatac atacgtcaca aaaagtggat cgcttattgt
     1021 ggtcgcaaga ctttactttt cacccactcc accatttctc aggtatgatg aggacgtcca
     1081 tgaccgaatt tggattccat tcttagataa caaaaattcc ttgttaagca cggaactctc
     1141 tgttgataca agtaacttct acaatgtgcc tcaaactgta gcgaaaactg ctgctgtccc
     1201 tttaaatgct actcagcctc tgaaaataaa ttggagtctc gacgacatca cttcacagtc
     1261 atatatatac atgcatttcg ctgaaatcga gaatcttgaa gctaatgaga ccagagaatt
     1321 caatattact tacaatggtg gcgaaaattg gttctcctat tttagacctc caaagtttcg
     1381 tataacaact gtatacaatc cagcagctgt gagttctcta gatgggaatt tcaacttcac
     1441 tttctcgatg accggtaact ctactcatcc tcctcttatc aacggccttg agatttatca
     1501 agttctagag cttccacagc ttgatacata ccaagatgaa gtttccgcta tgatgaatat
     1561 caagacaata tatggattga gcaaaaggtc tagctggcaa ggagatccat gtgctcctga
     1621 gttatataga tgggaaggtt taaactgtag ttatccaaac tttgcgccac cgcagatcat
     1681 atccttgaac ttgagtggaa gcaatttgag tggtaccata acatctgata tatccaagct
     1741 aacacatttg agagaactag atttatcaaa caatgactta tcaggagata ttccatttgt
     1801 tttttctgat atgaagaatt tgacactcat aaacttgagt ggaaacaaga atctaaatcg
     1861 ctcagttcca gagactcttc agaagaggat agataacaaa tctttaacac taattagaga
     1921 tgaaaccgga aaaaatagtc tggaggtgct gttccaggga ccagggagcg acaaaactca
     1981 cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc
     2041 cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt
     2101 ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt
     2161 gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag
     2221 cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc
     2281 caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg
     2341 agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag
     2401 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     2461 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     2521 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     2581 atgctccgtg atgcatgagg gtctgcacaa ccactacacg cagaagagcc tctccctgtc
     2641 tccgggtaaa tctagagggc ccttcgaagg taagcctatc cctaaccctc tcctcggtct
     2701 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     2761 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     2821 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2881 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2941 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3001 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     3061 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3121 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3181 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3241 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3301 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3361 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3421 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3481 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3541 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3601 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3661 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3721 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3781 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3841 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3901 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3961 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4021 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4081 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4141 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4201 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4261 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4321 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4381 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4441 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4501 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4561 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4621 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4681 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4741 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4801 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4861 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4921 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4981 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5041 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5101 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5161 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5221 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5281 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5341 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5401 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5461 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5521 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     5581 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5641 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
