LOCUS       sequence-297048-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G49770 X027_pECIA2 should be used with prey vector 
            AT5G49770 X027_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  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 5867)
  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: AT5G49770 X027_pECIA2
FEATURES             Location/Qualifiers
     source          1..5867
                     /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             2120..2143
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2849..2890
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2900..2917
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2940..2957)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2990..3124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3014..3033)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3068..3087
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3268..3284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3268..3284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3281..3303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3292..3308
                     /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(3316..3322,3323..3340,3341..3346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3361..3382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3499..3516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3670..4258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3750..3769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4429..5220,5221..5289))
                     /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     5052..5071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5290..5394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5462..5480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5518..5540)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5640..5659
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5853..5867,1..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 acagatggtt cagattttac
      541 tgctttacaa gccctgaaga acgaatggga cacgttatcc aagagttgga aaagctctga
      601 tccttgtgga accgaatggg ttggaattac atgtaacaac gataaccgtg ttgtttcaat
      661 atcattaacc aaccgtaact tgaaaggaaa gcttcctacc gaaatttcga cattgtctga
      721 attgcaaacc ttggatttga ctggcaatcc tgaattgtct ggaccgcttc cagcaaatat
      781 cggtaacctc aggaagttga ctttcttgag ccttatggga tgtgctttta atggtccaat
      841 ccctgattcc atagggaatc ttgaacaact tacaagactc tctctgaatt taaataaatt
      901 tagtggaaca attccggctt ccatgggacg gttatcgaaa ctatattggt tcgacatagc
      961 tgacaatcag ttagaaggaa agcttccagt ttctgatggg gcttctttac ctggacttga
     1021 catgcttctt caaactgggc attttcattt tggtaataac aagctttcag gcgaaatccc
     1081 agagaagctc tttagctcag aaatgacttt gctacatgtg ctgtttgatg gaaaccaatt
     1141 caccggtagc atcccagaga gcctcggcct agttcaaaat ttgactgtgt tacgtcttga
     1201 taggaataga ctaagtggag acattccttc aagtcttaat aatctcacaa atcttcaaga
     1261 actgcatttg tccgacaaca aatttacagg tagtcttcca aatttgacca gcttgaccag
     1321 tctctacaca ttggatgtga gcaataaccc gttggcgtta tcaccggttc catcatggat
     1381 ccctttctta aactctttgt caacattaag gttggaagat atccaacttg atggtccagt
     1441 accaacctcc ctttttagcc ctcttcaatt gcaaactgtt tccctaaagc acaatcttat
     1501 aaacacaaca ttggaccttg gtaccaacta tagcaaacaa ttggattttg tggatttacg
     1561 agacaacttc ataactggtt ataaatcacc agctaataac cccgtcaatg taatgttggc
     1621 agataatcaa gtgtgccaag acccggcgaa tcagctgagt ggctactgca atgcagtcca
     1681 accaaattcc acattttcta ccctaacaaa atgtggtaat cattgtggta aaggtaagga
     1741 accgaatcaa ggctgccact gtgtgtatcc acttacagga gtattcacct taaggtctcc
     1801 ttcattctcc ggattttcaa acaacagtaa cttcctaaag tttggggaga gtttaatgac
     1861 tttctttaag aatggcaagt atcctgtaga ctctgtggcc atgagaaaca taagcgagaa
     1921 tccaactgat tatcatcttt tgataaatct cttgatcttt ccatcgggac gcgaccgctt
     1981 taatcagacg gaaatggact ctattaattc tgcatttacc attcaggatt ataagcctcc
     2041 tccaagattt ggtccttata ttttcgtcgc cgatcagtac aaaacattct ctgatttaga
     2101 agattccaag acagtaagtc tggaggtgct gttccaggga ccagggagcg acaaaactca
     2161 cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc
     2221 cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt
     2281 ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt
     2341 gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag
     2401 cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc
     2461 caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg
     2521 agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag
     2581 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     2641 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     2701 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     2761 atgctccgtg atgcatgagg gtctgcacaa ccactacacg cagaagagcc tctccctgtc
     2821 tccgggtaaa tctagagggc ccttcgaagg taagcctatc cctaaccctc tcctcggtct
     2881 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     2941 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccaaacat
     3001 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3061 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3121 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3181 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     3241 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3301 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3361 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3421 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3481 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4081 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4321 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4501 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4561 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4621 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4681 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4741 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4801 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4861 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4921 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4981 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5041 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5101 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5161 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5221 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5281 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5341 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5401 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5461 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5521 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5581 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5641 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5701 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     5761 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5821 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
