LOCUS       sequence-296765-        4877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G45780 I3_pECIA2 should be used with prey vector 
            AT5G45780 I3_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4877)
  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 4877)
  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: AT5G45780 I3_pECIA2
FEATURES             Location/Qualifiers
     source          1..4877
                     /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             1130..1153
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1859..1900
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1910..1927
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1950..1967)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2000..2134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2024..2043)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2078..2097
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2278..2294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2278..2294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2291..2313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2302..2318
                     /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(2326..2332,2333..2350,2351..2356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2371..2392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2509..2526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2680..3268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2760..2779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3439..4230,4231..4299))
                     /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     4062..4081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4300..4404)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4472..4490
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4528..4550)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4650..4669
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4863..4877,1..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 tctcccaagg gtgttaacta
      541 tgaagtggct gcgttaatgt cagtgaagaa caagatgaaa gatgagaaag aggttttgtc
      601 tggttgggat attaactctg ttgatccttg tacttggaac atggttggtt gttcttctga
      661 aggttttgtg gtttctctag agatggctag taaaggatta tcagggatac tatctactag
      721 tattggggaa ttaactcatc ttcatacttt gttacttcag aataatcagt taactggtcc
      781 gattccttct gagttaggcc aactctctga gcttgaaacg cttgatttat cggggaatcg
      841 gtttagtggt gaaatcccag cttctttagg gttcttaact cacttaaact acttgcggct
      901 tagcaggaat cttttatctg ggcaagtccc tcacctcgtc gctggcctct caggtctttc
      961 tttcttggat ctatctttca acaatctaag cggaccaact ccgaatatat cagcaaaaga
     1021 ttacaggatt gtaggaaatg catttctttg tggtccagct tcccaagagc tttgctcaga
     1081 tgctacacct gtgagaaatg cgacgggttt gtctgaaaag gacaatagcc tggaggtgct
     1141 gttccaggga ccagggagcg acaaaactca cacatgccca ccgtgcccag cacctgaact
     1201 cctgggggga ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc
     1261 ccggacccct gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa
     1321 gttcaactgg tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga
     1381 gcagtacaac agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct
     1441 gaatggcaag gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa
     1501 aaccatctcc aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc
     1561 ccgggaggag atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc
     1621 cagcgacatc gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac
     1681 gcctcccgtg ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa
     1741 gagcaggtgg cagcagggga acgtcttctc atgctccgtg atgcatgagg gtctgcacaa
     1801 ccactacacg cagaagagcc tctccctgtc tccgggtaaa tctagagggc ccttcgaagg
     1861 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca
     1921 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     1981 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     2041 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2101 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2161 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2221 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     2281 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2341 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2401 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     2461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4441 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4501 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4561 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4621 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4681 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     4741 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4801 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     4861 ttcccagtca cgacgtt
//
