LOCUS       sequence-297013-        5738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G62950 X004_pECIA2 should be used with prey vector 
            AT1G62950 X004_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5738)
  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 5738)
  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: AT1G62950 X004_pECIA2
FEATURES             Location/Qualifiers
     source          1..5738
                     /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             1991..2014
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2720..2761
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2771..2788
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2811..2828)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2861..2995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2885..2904)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2939..2958
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3139..3155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3139..3155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3152..3174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3163..3179
                     /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(3187..3193,3194..3211,3212..3217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3232..3253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3370..3387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3541..4129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3621..3640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4300..5091,5092..5160))
                     /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     4923..4942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5161..5265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5333..5351
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5389..5411)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5511..5530
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5724..5738,1..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 tcggattcta ttattaccga
      541 acgagagatt ctacttcaat tcaaagacaa catcaacgat gatccgtaca acagtttagc
      601 ttcttgggtc tctaatgctg acctctgtaa cagcttcaat ggtgtatctt gtaaccaaga
      661 aggattcgtc gaaaagatcg ttctttggaa cactagtctc gccggaaccc taacgccggc
      721 attgtctggt ttaacttctt taagggtttt gactctgttt ggcaacagga ttacaggtaa
      781 tttacctttg gattacttaa agttacagac tttatggaag atcaatgtta gttccaatgc
      841 attgtctggt ttggttcctg aatttatcgg tgatttgcct aatttaaggt tcttggattt
      901 gtcgaaaaat gcgttctttg gagagattcc aaattctctg tttaaattct gttacaagac
      961 aaagtttgtt tcattgtctc ataacaatct ctcaggatca atccctgaat caatcgtaaa
     1021 ctgtaacaat ctcattggtt tcgatttctc ttacaatggc attaccggtt tgttgcctcg
     1081 gatctgcgat attccggttc ttgagtttgt atctgttaga agaaatttgt tgtctggtga
     1141 tgtgtttgag gagatatcga agtgtaagag attgagtcat gtcgatatag gaagcaattc
     1201 tttcgatgga gtagcgtctt tcgaggttat tggattcaag aacctgactt attttaatgt
     1261 gtctggaaac cggtttagag gggagattgg tgagattgtt gattgtagtg aaagtttgga
     1321 gtttttggat gcttcttcga atgagttaac tggtaatgta cctagcggta taaccggatg
     1381 caaaagtctt aagcttttgg acttggaatc gaataggctc aatgggagtg tacctgtagg
     1441 tatggggaag atggagaagc tctctgtgat caggttaggc gataacttta tagatgggaa
     1501 gctaccactt gagctcggaa atcttgagta tctacaggtt ttgaatttgc ataatctcaa
     1561 tctcgttggt gaaataccag aggatttatc aaactgcaga ctacttcttg aactagatgt
     1621 ttctggaaat ggtttggaag gagagatccc taagaatcta ttgaacttaa caaacttaga
     1681 gattcttgat ctgcatcgga accgaatcag tggaaacatt ccgcctaatc tcgggagcct
     1741 ctcgagaatc cagttcctcg atctttcaga gaatttgctt tctggtccta tcccgtcttc
     1801 cctcgagaat ttgaaaagac tgactcattt caatgtttct tacaacaacc tctccggcat
     1861 tatcccaaag attcaagctt cgggagcttc ttcgttttcc aacaaccctt tcctctgcgg
     1921 tgatcctctt gaaacaccgt gcaatgctct cagaaccgga tcaagatcaa gaaaaaccaa
     1981 agctctaagc ctggaggtgc tgttccaggg accagggagc gacaaaactc acacatgccc
     2041 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     2101 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     2161 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     2221 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     2281 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     2341 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     2401 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     2461 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     2521 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     2581 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     2641 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     2701 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2761 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2821 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     2881 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2941 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3001 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3061 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     3121 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3181 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3241 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3301 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3361 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3961 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4201 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4261 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4321 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4381 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4441 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4501 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4561 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4621 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4681 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4741 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4801 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4861 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4921 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4981 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5041 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5101 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5161 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5221 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5281 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5341 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5401 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5461 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5521 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5581 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5641 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5701 aagttgggta acgccagggt tttcccagtc acgacgtt
//
