LOCUS       sequence-298970-        6119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT3G47580 L8_pECIA2 should be used with prey vector 
            AT3G47580 L8_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6119)
  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 6119)
  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: AT3G47580 L8_pECIA2
FEATURES             Location/Qualifiers
     source          1..6119
                     /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             2372..2395
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3101..3142
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3152..3169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3192..3209)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3242..3376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3266..3285)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3320..3339
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3520..3536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3520..3536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3533..3555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3544..3560
                     /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(3568..3574,3575..3592,3593..3598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3613..3634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3751..3768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3922..4510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4002..4021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4681..5472,5473..5541))
                     /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     5304..5323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5542..5646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5714..5732
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5770..5792)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5892..5911
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6105..6119,1..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 accgatgaaa ctgataggca
      541 agcattgttg gagttcaagt ctcaagtttc tgaaggcaaa agagatgtct tatcctcatg
      601 gaataactca ttccctctat gcaactggaa gtgggttaca tgtggtcgta aacacaagag
      661 agttactcat ttgaacctcg ggggattaca gttgggcggg atagtatcac catctatcgg
      721 taatgtatcg tttctcatat cacttgatct gtctgataac gcttttgggg gtatcatccc
      781 tcgagaggtg ggaaacttgt ttagacttga acacttgtat atggccttca attctctaga
      841 gggagggatt ccagccactc tatctaactg ctctagattg ttgaaccttg atttatattc
      901 aaatcctctt agacaaggtg ttccttctga actagggtca ttgacgaaac tcgttatttt
      961 agatcttggt cgaaacaacc tgaaaggcaa gctccctaga tctctaggaa acttgacctc
     1021 tttaaaatcg cttggcttca cagataacaa tatagaagga gaagttccag atgaattagc
     1081 tagactgagt caaatggtgg gtcttgggtt atcaatgaac aaattctttg gcgtttttcc
     1141 tcccgccatc tacaatttgt ccgcacttga ggacttattc ctctttggta gtggcttctc
     1201 gggtagccta aagcctgatt ttggtaatct gctaccaaac attcgtgagt taaatctggg
     1261 agaaaatgat ctagtgggag ccattccaac aacactttcc aatatttcaa ctctacaaaa
     1321 gtttggaatc aacaaaaaca tgatgacagg aggtatatat ccgaactttg gaaaagtacc
     1381 gagtttgcaa tacctagatc ttagtgagaa tcctctagga agttacactt ttggagatct
     1441 tgaatttatt gattctttga ctaattgcac ccatttgcaa ctcttaagtg ttggttacac
     1501 taggcttggg ggtgccttgc ctacatccat tgccaatatg tccacagaac tcatcagttt
     1561 aaaccttata ggaaaccact tttttggaag cattcctcaa gatattggga atctcatagg
     1621 cctacaaaga cttcagttgg gaaaaaatat gttgaccgga ccactcccaa cttctttggg
     1681 aaagcttcta agattggggt tattaagtct atattcaaat aggatgtcag gagagattcc
     1741 atcttttata ggcaacctaa ctcaattaga aatactatat ttgtccaaca atagttttga
     1801 gggaattgtt cctccgagtc tcgggaagtg tagtcacatg cttgatttac gcattgggta
     1861 taacaagttg aatgggacta taccgaagga aattatgcaa atcccaaccc ttgttaacct
     1921 aagcatggaa ggtaactctt tgagcggctc tctaccaaat gatattggaa gtctacaaaa
     1981 tcttgtaaaa ctatcacttg agaacaataa gttttcagga catctcccac aaaccttggg
     2041 aaattgtctc gcgatggaac aactttttct acaaggaaat tcttttgatg gagcaattcc
     2101 aaatataaga gggttgatgg gtgtaagaag agttgatttg tcaaacaatg atctatcggg
     2161 aagtatacct gaatattttg caaacttctc caagttggaa tatctcaatc tatctattaa
     2221 caattttacg ggaaaggtgc cttccaaagg aaattttcag aactctacaa tagtttttgt
     2281 atttggaaac aaaaacctat gtggaggcat caaggatctg aaactaaagc catgcctcgc
     2341 ccaagaacca ccagtggaga caaagcattc cctggaggtg ctgttccagg gaccagggag
     2401 cgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg gaccgtcagt
     2461 cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc ctgaggtcac
     2521 atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact ggtacgtgga
     2581 cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca acagcacgta
     2641 ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca aggagtacaa
     2701 gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct ccaaagccaa
     2761 agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggagg agatgaccaa
     2821 gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca tcgccgtgga
     2881 gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg tgctggactc
     2941 cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt ggcagcaggg
     3001 gaacgtcttc tcatgctccg tgatgcatga gggtctgcac aaccactaca cgcagaagag
     3061 cctctccctg tctccgggta aatctagagg gcccttcgaa ggtaagccta tccctaaccc
     3121 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     3181 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     3241 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3301 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3361 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3421 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     3481 tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4741 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4861 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5101 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5341 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5521 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5581 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5641 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5701 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5761 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5821 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5881 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5941 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     6001 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6061 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
