LOCUS       sequence-296647-        5717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G16900 O2_pECIA2 should be used with prey vector 
            AT5G16900 O2_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5717)
  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 5717)
  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: AT5G16900 O2_pECIA2
FEATURES             Location/Qualifiers
     source          1..5717
                     /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             1970..1993
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2699..2740
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2750..2767
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2790..2807)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2840..2974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2864..2883)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2918..2937
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3118..3134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3118..3134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3131..3153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3142..3158
                     /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(3166..3172,3173..3190,3191..3196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3211..3232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3349..3366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3520..4108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3600..3619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4279..5070,5071..5139))
                     /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     4902..4921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5140..5244)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5312..5330
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5368..5390)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5490..5509
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5703..5717,1..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 agtttggatt gcgggttgcc
      541 gagtaacgag cctccatata ttgaaccggt aaccggatta gtattctcat cggatgcgga
      601 tcacatccct agtggaataa gcggtagaat ccagaagaat ctggaagcag tacacataaa
      661 gccgtatttg tttctgagat actttccaga tgggctacga aattgctata ctctagatgt
      721 ccttcagaac cgaagataca tgatcaaagc tgtgtttgtg tatggaaatt acgatggtta
      781 taatgattat ccgagttttg atctgtatct aggtcctaat aagtgggtaa gagttgactt
      841 ggaaggaaag gtcaatggca gtgttgaaga gattattcac ataccaagtt caaactctct
      901 gcagatttgt cttgtcaaga caggtaattc tttgccgttc atatccgccc tggaactgag
      961 gcttttgaga aacgatactt atgttgtcca agacgtttcc ttgaagcatt tatttcggag
     1021 gtactataga cagtcagatc ggttaatccg gtatcctgac gatgtctatg atcgtgtttg
     1081 gtccccattc ttcttaccgg aatggacgca gataactacc agtttagatg tgaataactc
     1141 taacaactac gagccaccaa aagcagcatt aacatcagct gccacacctg gagacaatgg
     1201 aacgcgattg accattattt ggactttgga taaccctgat gagcaaattc acctgtatgt
     1261 tcactttgcc gagcttgaac ctgttgggga aaacactgac gaggccttga gaactttatt
     1321 tacaaggacc ttttactttg tggtgaatgg aaagattagt tatgatgaga gtattactcc
     1381 tttagacttg gctgtaagta ctgtagagac tgtggtgaac aaatgcgatg gcggtaactg
     1441 cagtttgcag ctggtacgca gcgaagcgtc tccgggagta cgtgtacctc tggttaatgc
     1501 tatggaggca ttcactgcaa tcaagtttcc acattctgaa acaaatcctg atgatgttat
     1561 atctatcaag gttatccaag ctacctatga actgagcaga gtcgactggc aaggagatcc
     1621 gtgtcttccc cagcagtttt tgtggactgg cctaaactgc agctacatga acatgtctac
     1681 gtcaccaaga atcatttcct tagacctgtc ctcacataaa ttaactggga aaatagtgcc
     1741 agatatacaa aatctgaccc agctacagaa actggacttg tcgaataaca agttgaccgg
     1801 aggtgtgcct gagtttctag ccaacatgaa gtcactgttg ttcataaacc taagcaataa
     1861 caatcttgtt ggttccattc ctcaagcgct tcttgaccga aaaaatctga agttggagtt
     1921 tgaaggaaat ccaaagcttt gcgccacagg tccgtgtaac agttcaagtc tggaggtgct
     1981 gttccaggga ccagggagcg acaaaactca cacatgccca ccgtgcccag cacctgaact
     2041 cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc
     2101 ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa
     2161 gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga
     2221 gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct
     2281 gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa
     2341 aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc
     2401 ccgggaggag atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc
     2461 cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac
     2521 gcctcccgtg ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa
     2581 gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcatgagg gtctgcacaa
     2641 ccactacacg cagaagagcc tctccctgtc tccgggtaaa tctagagggc ccttcgaagg
     2701 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     2761 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     2821 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     2881 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2941 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3001 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3061 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     3121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3661 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5281 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5341 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5401 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5461 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5521 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     5581 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5641 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5701 ttcccagtca cgacgtt
//
