LOCUS       sequence-227478-        4868 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bait vector AT3G28450 X043_pECIA2 should be used with prey vector 
            AT3G28450 X043_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4868)
  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 4868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AT3G28450 X043_pECIA2
FEATURES             Location/Qualifiers
     source          1..4868
                     /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             1121..1144
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1850..1891
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1901..1918
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1941..1958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1991..2125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2015..2034)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2069..2088
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2269..2285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2269..2285)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2282..2304)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2293..2309
                     /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(2317..2323,2324..2341,2342..2347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2362..2383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2500..2517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2671..3259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2751..2770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3430..4221,4222..4290))
                     /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     4053..4072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4291..4395)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4463..4481
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4519..4541)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4641..4660
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4854..4868,1..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 tctgttatgg ctgcagatga
      541 agacgacata agatgcttaa gaggactcaa agcatctctc acagatcctc aaaacgcgtt
      601 gaaatcatgg aacttcgaca acacaactct tgggtttctc tgtaacttcg ttggtgtgtc
      661 ttgttggaac aatcaggaga atagggttat caatcttgag cttcgtgata tgggtttatc
      721 tggtaaaatc ccagattctc ttcaatactg tgcgagttta caaaaacttg atctttctag
      781 taatcgattg tctggtaata tccctacaga gttgtgtaat tggttacctt ttttggtatc
      841 tcttgatttg tcaaacaatg aattgaatgg tgagattcct cctgatttag ctaagtgtag
      901 ctttgtgaat tctttggttt tgtctgataa ccggctttcg ggtcaaatcc cggttcagtt
      961 ctcggcttta gggaggttag ggaggttctc tgttgctaat aatgatctct caggtcgcat
     1021 tcctgtgttc tttagctcac cgagttattc ctctgatgat tttagtggga ataaaggtct
     1081 ttgtggtcgt cctttatctt cgagttgtgg tggtttaagt ctggaggtgc tgttccaggg
     1141 accagggagc gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg
     1201 accgtcagtc ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc
     1261 tgaggtcaca tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg
     1321 gtacgtggac ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa
     1381 cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
     1441 ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc
     1501 caaagccaaa gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga
     1561 gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat
     1621 cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt
     1681 gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
     1741 gcagcagggg aacgtcttct catgctccgt gatgcatgag ggtctgcaca accactacac
     1801 gcagaagagc ctctccctgt ctccgggtaa atctagaggg cccttcgaag gtaagcctat
     1861 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg
     1921 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     1981 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2041 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2101 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     2161 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     2221 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     2281 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2341 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2401 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2461 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2521 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2581 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2641 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2701 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2761 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2821 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2881 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2941 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3001 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3061 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3121 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3181 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3241 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3301 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3361 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3421 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3481 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3541 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3601 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3661 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3721 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3781 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3841 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3901 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3961 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4021 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4081 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4141 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4201 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4261 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4321 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4381 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4441 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     4501 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     4561 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4621 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     4681 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     4741 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     4801 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     4861 acgacgtt
//
