LOCUS       sequence-238709-        6119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector AT1G08590 X046_pECIA2 should be used with prey vector 
            AT1G08590 X046_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: AT1G08590 X046_pECIA2
FEATURES             Location/Qualifiers
     source          1..6119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     194..210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        216..642
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     660..713
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2565..2588
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3294..3335
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3345..3362
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3385..3402)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3435..3569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3459..3478)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3513..3532
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3713..3729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3713..3729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3726..3748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3737..3753
                     /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(3761..3767,3768..3785,3786..3791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3806..3827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3944..3961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4115..4703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4195..4214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4874..5665,5666..5734))
                     /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     5497..5516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5735..5839)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5907..5925
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5963..5985)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6085..6104
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
       61 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      121 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      181 cagtcacgac gttgtaaaac gacggccagt gaattaattc gttgcaggac aggatgtggt
      241 gcccgatgtg actagctctt tgctgcaggc cgtcctatcc tctggttccg ataagagacc
      301 cagaactccg gccccccacc gcccaccgcc acccccatac atatgtggta cgcaagtaag
      361 agtgcctgcg catgccccat gtgccccacc aagagctttg catcccatac aagtccccaa
      421 agtggagaac cgaaccaatt cttcgcgggc agaacaaaag cttctgcaca cgtctccact
      481 cgaatttgga gccggccggc gtgtgcaaaa gaggtgaatc gaacgaaaga cccgtgtgta
      541 aagccgcgtt tccaaaatgt ataaaaccga gagcatctgg ccaatgtgca tcagttgtgg
      601 tcagcagcaa aatcaagtga atcatctcag tgcaactaaa ggggggatcc gatctcaata
      661 tgaagttatg catattactg gccgtcgtgg cctttgttgg cctctcgctc gggtctgaaa
      721 catttcaaaa ctccgaacaa gagatcttgt tggccttcaa gtcagacctt tttgatcctt
      781 caaataatct ccaagattgg aagagacctg agaatgccac aacattttct gagttggttc
      841 attgtcattg gacaggagtt cactgtgatg caaatggcta tgtggctaag cttcttctct
      901 ccaacatgaa tctcagcgga aacgtgtcag atcagattca gagcttccct tctctgcaag
      961 cccttgatct cagcaataac gcatttgagt cttctcttcc taaatctctt tctaatctca
     1021 cgtctttgaa ggtcattgat gtgagtgtga acagcttctt tggaaccttc ccttatggtc
     1081 taggaatggc aactggtctt acacacgtca atgcctcaag caacaatttt tcggggtttc
     1141 tacctgaaga tcttggcaac gcaactacac ttgaggtttt ggactttcga ggaggttact
     1201 tcgagggctc agtgccttct tctttcaaga atctcaagaa tctgaagttt ctcggtcttt
     1261 cgggtaacaa ctttggtgga aaggtaccga aggtgattgg tgagctctct tccttggaga
     1321 ccattattct tggttacaat ggcttcatgg gtgagattcc agaagagttt gggaaattga
     1381 cgcgccttca gtaccttgat ttggctgttg gaaaccttac aggccagatt ccatcttctt
     1441 tggggcagct gaaacaactt acaacagttt atctctacca gaaccgtctc acagggaaac
     1501 ttcctcgaga acttggtggt atgacttctc ttgtgtttct tgatctgtct gataaccaga
     1561 tcacaggaga aatacctatg gaagtaggtg agttgaagaa tctgcagctt ctaaatttga
     1621 tgaggaatca actcacggga atcattccgt caaaaattgc tgaattacca aaccttgagg
     1681 ttcttgagct gtggcagaac tctctcatgg gttcattacc ggtgcatctc ggcaaaaatt
     1741 ccccattgaa gtggttagac gtctcatcaa acaagctttc aggagatata ccatcaggat
     1801 tgtgctattc cagaaacctc acaaagctca ttctcttcaa caactcattc tcaggtcaga
     1861 ttccggagga aatcttctca tgtccgactt tagttcgtgt tcgaatccag aaaaatcaca
     1921 tttccggatc tataccagct ggttcaggtg acttaccaat gcttcagcat ctagagctcg
     1981 caaaaaacaa cctcaccggt aaaattccag atgatatagc cttatccaca tcactctcat
     2041 ttatagacat atcatttaat catctctcat ccctatcttc ctccattttc tcaagtccta
     2101 atcttcaaac cttcattgcc tcacataaca actttgccgg aaaaattcct aaccagattc
     2161 aagatcgtcc atctttgtct gtcctcgatc tttccttcaa ccatttctca ggtggaatcc
     2221 cagaaaggat cgcttctttt gaaaaacttg tgagtctaaa tctcaagagc aaccaattgg
     2281 ttggagaaat ccctaaggca ttagccggga tgcacatgtt agctgttctt gatctctcaa
     2341 acaactcttt gacggggaat ataccagcgg acttgggagc ttctccaaca ctggagatgt
     2401 taaacgtgtc attcaacaaa ctcgacggac ccattccttc aaacatgctg tttgcagcca
     2461 tagacccaaa agacctcgtg ggaaacaatg gcttatgcgg tggagttcta ccaccttgct
     2521 ctaagagcct agctttgtca gcaaaaggcc gaaatccagg gagactggag gtgctgttcc
     2581 agggaccagg gagcgacaaa actcacacat gcccaccgtg cccagcacct gaactcctgg
     2641 ggggaccgtc agtcttcctc ttccccccaa aacccaagga caccctcatg atctcccgga
     2701 cccctgaggt cacatgcgtg gtggtggacg tgagccacga agaccctgag gtcaagttca
     2761 actggtacgt ggacggcgtg gaggtgcata atgccaagac aaagccgcgg gaggagcagt
     2821 acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg
     2881 gcaaggagta caagtgcaag gtctccaaca aagccctccc agcccccatc gagaaaacca
     2941 tctccaaagc caaagggcag ccccgagaac cacaggtgta caccctgccc ccatcccggg
     3001 aggagatgac caagaaccag gtcagcctga cctgcctggt caaaggcttc tatcccagcg
     3061 acatcgccgt ggagtgggag agcaatgggc agccggagaa caactacaag accacgcctc
     3121 ccgtgctgga ctccgacggc tccttcttcc tctacagcaa gctcaccgtg gacaagagca
     3181 ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca tgagggtctg cacaaccact
     3241 acacgcagaa gagcctctcc ctgtctccgg gtaaatctag agggcccttc gaaggtaagc
     3301 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     3361 attgagttta aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat
     3421 cagcctcgat cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     3481 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3541 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3601 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3661 tggctgatta tgatcagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
     3721 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3781 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3841 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3901 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3961 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4021 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4081 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4141 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4201 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4261 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4321 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4381 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4441 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4501 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4561 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4621 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4681 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4861 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4921 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4981 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5041 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5101 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5161 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5221 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5281 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5341 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5401 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5461 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5521 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5581 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5641 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5701 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5761 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5821 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5881 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5941 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     6001 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6061 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccat
//
