LOCUS       sequence-296652-        5678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT5G59670 O6_pECIA2 should be used with prey vector 
            AT5G59670 O6_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5678)
  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 5678)
  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: AT5G59670 O6_pECIA2
FEATURES             Location/Qualifiers
     source          1..5678
                     /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             1931..1954
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2660..2701
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2711..2728
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2751..2768)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2801..2935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2825..2844)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2879..2898
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3079..3095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3079..3095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3092..3114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3103..3119
                     /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(3127..3133,3134..3151,3152..3157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3172..3193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3310..3327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3481..4069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3561..3580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4240..5031,5032..5100))
                     /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     4863..4882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5101..5205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5273..5291
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5329..5351)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5451..5470
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5664..5678,1..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 gcgggctacc
      541 cgcaaatgag acgtctcctt atacagagac tcaaactggt ctgttgttct cttcggacgc
      601 aacattcatc cagagcggaa aaactggtag agtccaagca aatcaagaga gtaaattctt
      661 gaagccatac agaacgctga gatatttccc agagggagta cggaattgct acaatctgag
      721 tgtgttcaag gagcgtaaat atttgatcgc tgctagtttt ttatatggaa attacgatgg
      781 tcataacata gcccctgtgt ttgatcttta tcttggacca aatttatggg cgaaaataga
      841 tttacaagat gtgaatggta caggggaaga gatcttacac atcccaacat cgaactcgtt
      901 gcagatatgt cttgttcaga ctggggaaac tacaccgctg atatcctctt tggaattacg
      961 gccaatgaga actggaagct ataccactgt gtctggttcc ctgaaaacat acaggcgttt
     1021 atattttaaa aagtctggat ctaggctacg gtacagtaaa gatgtttatg atcgctcatg
     1081 gtttccgcgc ttcatggacg aatggaccca gatttccact gctctcgggg tgattaatac
     1141 caacatctac caaccaccgg aagatgcgct gaaaaatgct gccacgccta ctgatgctag
     1201 tgcgccactg acatttaaat ggaattcaga aaaattggat gtccaatatt acttttacgc
     1261 acactatgct gagatccaag acttgcaggc aaatgatacc agagaattca acatcttatt
     1321 gaatggacaa aatttatctg tgactggccc tgaagtccca gataagctaa gtataaagac
     1381 cttccaaagc tcgtcaccga taagttgcaa tggatgggca tgtaattttc agctgataag
     1441 aaccaaaaga tcaactcttc cacctctact taatgctttg gaagtctaca cagttattca
     1501 gtttccacgg tccgaaacag atgaaagcga cgtggttgct atgaaaaaca tctcagccag
     1561 ctatggattg agtagaatca actggcaagg cgatccatgc ttccctcagc agcttaggtg
     1621 ggacgcttta gactgcacca acagaaatat ctcccaacca ccaagaatca cttctttaaa
     1681 cttgtcttca agtcgtttaa atggaaccat agcagctgcc attcaaagta ttacacaact
     1741 tgaaacactg gacttgtcgt ataataactt aactggagag gtgccagagt ttcttggcaa
     1801 aatgaaatcg ttgtcggtca taaacttaag cgggaacaat cttaatggtt caattccgca
     1861 agctcttcga aagaaaagac taaaactata ccttgaagga aacccaaggc ttataaaacc
     1921 acctaagaag ctggaggtgc tgttccaggg accagggagc gacaaaactc acacatgccc
     1981 accgtgccca gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc
     2041 caaggacacc ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag
     2101 ccacgaagac cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc
     2161 caagacaaag ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac
     2221 cgtcctgcac caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc
     2281 cctcccagcc cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca
     2341 ggtgtacacc ctgcccccat cccgggagga gatgaccaag aaccaggtca gcctgacctg
     2401 cctggtcaaa ggcttctatc ccagcgacat cgccgtggag tgggagagca atgggcagcc
     2461 ggagaacaac tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta
     2521 cagcaagctc accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt
     2581 gatgcatgag ggtctgcaca accactacac gcagaagagc ctctccctgt ctccgggtaa
     2641 atctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2701 gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2761 gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata
     2821 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2881 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2941 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     3001 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc
     3061 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3121 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3181 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3241 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3901 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4141 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4201 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4261 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4321 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4381 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4441 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4501 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4561 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4621 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4681 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4741 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4801 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4861 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4921 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4981 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5041 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5101 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5161 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5221 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5281 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5341 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5401 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5461 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5521 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5581 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5641 aagttgggta acgccagggt tttcccagtc acgacgtt
//
