LOCUS       sequence-298764-        5978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G34420 X003_pECIA2 should be used with prey vector 
            AT1G34420 X003_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  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 5978)
  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: AT1G34420 X003_pECIA2
FEATURES             Location/Qualifiers
     source          1..5978
                     /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             2231..2254
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2960..3001
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3011..3028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3051..3068)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3101..3235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3125..3144)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3179..3198
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3379..3395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3379..3395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3392..3414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3403..3419
                     /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(3427..3433,3434..3451,3452..3457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3472..3493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3610..3627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3781..4369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3861..3880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4540..5331,5332..5400))
                     /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     5163..5182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5401..5505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5573..5591
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5629..5651)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5751..5770
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5964..5978,1..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 tcgttgaacc agacaaacac
      541 aatgatcgaa ctctctagtt tccttaatat ctcggattgg aatcttcctg gttctgagag
      601 gaatccatgt tcatggaacg gtgttctctg cagcttacca gacaatagct ctgtgattag
      661 cctgtctctc tctaactttg atctctccaa ttcttccttc ttaccacttg tctgcaatct
      721 ccagactttg gagtctcttg atgtctccaa caacagattg agctcaatcc cagaagggtt
      781 cgttacaaat tgcgaaagac tcattgctct gaagcatctg aacttcagca cgaacaagtt
      841 ttctacttcc cctggtttcc gcggtttctc caaattggcg gttcttgatt tctctcacaa
      901 cgtgttgagt gggaacgttg gagactatgg ttttgatggg ttggttcagt tgaggagttt
      961 gaatcttagc ttcaacagat taactggttc tgttccggtt catttgacca aaagtctaga
     1021 gaagcttgag gtttcagata atagtttgag cggtactatt cccgaaggca tcaaagatta
     1081 tcaagaacta acgctgatag atctcagtga taatcagctt aacggctcta tcccgagttc
     1141 gttgggaaac ctctccaagt tagaaagttt acttctctcc aacaactatc ttagtggact
     1201 aatcccagaa tcactttcaa gcatccagac attgcgtcgg tttgcagcga accgtaaccg
     1261 attcacggga gaaattccat cagggctcac aaaacacctc gagaacttag atctcagttt
     1321 caacagttta gcggggtcga ttccaggtga ccttttgtcg cagcttaaac tcgtctctgt
     1381 ggatctgtca tctaatcaac ttgttggatg gataccacaa agcatttcgt ctagcctggt
     1441 taggctaaga ctaggaagca ataagctaac agggtcagta ccctctgttg cgtttgaatc
     1501 gctgcagctc ctaacgtatc tggagatgga caacaacagt ttaactggct tcatacctcc
     1561 ttcatttgga aatcttgtga gcttgaatct cctgaacttg gctatgaacg agttcaccgg
     1621 gatcttacct cctgcatttg gaaaccttag caggcttcaa gtgataaaac tacagcaaaa
     1681 caagcttacg ggagaaatac cagatacaat cgcatttttg agtaacctct tgattcttaa
     1741 catcagctgc aattctctaa gtggatccat accgccttcg ctttctcagc taaagaggct
     1801 ttccaacatg aacttgcaag gcaacaatct gaatggtacc atacctgata acattcaaaa
     1861 tctggaggat ttgatagaac ttcagctggg acaaaaccag ctacgaggga ggatcccagt
     1921 aatgccacga aagttgcaaa tttccttaaa tctcagctac aacttgtttg aaggatctat
     1981 ccctactaca ttgtctgaac ttgaccgctt ggaagttctt gatctgtcaa ataacaactt
     2041 ctccggcgaa atacctaatt tcctgagcag attaatgtcc cttacacagc tcatactctc
     2101 caacaatcag cttaccggaa atattccaag gttcacccat aatgtttcag tcgatgtcag
     2161 aggaaaccct ggggttaagt tgaaaactga gaacgaagtc tctatccaga gaaatccatc
     2221 agggaaaagc ctggaggtgc tgttccaggg accagggagc gacaaaactc acacatgccc
     2281 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     2341 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     2401 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     2461 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     2521 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     2581 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     2641 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     2701 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     2761 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     2821 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     2881 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     2941 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     3001 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     3061 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     3121 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3181 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3241 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3301 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     3361 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3421 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3481 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3541 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4141 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4201 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5521 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5581 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5641 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5701 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5761 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5821 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5881 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5941 aagttgggta acgccagggt tttcccagtc acgacgtt
//
