LOCUS       sequence-274537-        7037 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Prey vector AT1G56130 X071_pECIA14 should be used with bait vector 
            AT1G56130 X071_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  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 7037)
  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: AT1G56130 X071_pECIA14
FEATURES             Location/Qualifiers
     source          1..7037
                     /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             2333..2356
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4031..4054
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4070..4087
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4110..4127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4160..4294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4184..4203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4238..4257
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4438..4454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4438..4454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4451..4473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4462..4478
                     /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(4486..4492,4493..4510,4511..4516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4531..4552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4669..4686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4840..5428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4920..4939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5599..6390,6391..6459))
                     /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     6222..6241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6460..6564)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6632..6650
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6688..6710)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6810..6829
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(7023..7037,1..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 caaaccggag ccactactca
      541 tcccgacgaa gcgcgagctt tgaactcaat tttcgcggct tggaagatcc aggcgccgag
      601 agaatggaac atcagcggcg aactttgctc cggcgccgct atcgacgcca gtgtactcga
      661 cagcaaccct gcctacaatc ctctcatcaa atgcgactgt agtttccaaa actccacaat
      721 ctgccgcatc actaacatca aggtttatgc gatagatgtt gtaggaccta tccctccaga
      781 gctctggact ttaacatacc tcaccaatct gaacttgggt caaaatgttc tcactggctc
      841 acttcctcct gcaattggaa atctgactcg aatgcaatgg atgacttttg ggatcaatgc
      901 gttgtctggc cctgttccta aggaaattgg tttgcttaca gatttaagat tacttggtat
      961 tagttcaaat aacttttccg gttctatacc agatgagatt gggagatgta caaaactaca
     1021 acagatgtac atagatagtt ctggactcag cgggagaata cctttatcat ttgctaacct
     1081 cgtgcagctg gaacaagctt ggatcgcgga tctggaagtt acagatcaga taccagactt
     1141 tataggagat tggaccaaac ttactacctt gagaattatt ggaactggtc tgagtggtcc
     1201 gataccttcg tcattttcca acttgacttc gttgacagaa ttgaggcttg gtgatatatc
     1261 cagtggaagc tcttctcttg atttcatcaa agacatgaaa tctcttagtg tattagtttt
     1321 gaggaacaac aatctcactg ggacaatacc gtctactatt ggagaacact caagtttgcg
     1381 acaagttgat ttaagcttca acaaactaca tggaccaatt ccggcttcac ttttcaactt
     1441 aagtcagctt actcacttgt ttctgggaaa caacacgttg aatggctcct ttcccactca
     1501 aaagacgcag tctctgagaa atgtagatgt gtcgtacaat gatttgtctg gaagtcttcc
     1561 ttcatgggtc agcttaccaa gcttgaaact caacctagtt gctaacaact tcacattaga
     1621 aggtcttgac aacagggttt taccaggact gaactgcctg cagaaaaact tcccctgcaa
     1681 tcgaggcaaa ggaatctatt ctgacttttc aatcaactgc ggaggcccag agaaaagatc
     1741 tgtaactgga gcactatttg agagggagga tgaggatttt ggaccagctt catttttcgt
     1801 gagtgctggt cagagatggg cagctagtag tgtaggactt tttgccggaa gtagcaacaa
     1861 tatatacata gctacttcac aatcacaatt tgtcaacact ttggactcag agctatttca
     1921 atcagcaaga ctttctgcat cttccgtaag gtattatggg ttggggctag aaaatggagg
     1981 atatacagta acccttcagt ttgctgaaat acaaattcta ggttctactt ctaccacttg
     2041 gaaaggtttg ggacgacgac gttttgacat ttatgtccag ggaagacttg ttgaaaagga
     2101 ttttgatgta cgtagaacag ctggtgactc tactgttaga gcagttcaga gagtatataa
     2161 ggcaaatgta tcagaaaatc acctcgaagt tcatcttttc tgggctggga aaggaacatg
     2221 ttgtatacct atccaagggg cttatggacc attaatatca gcagtcagtg caacaccaga
     2281 tttcacacca actgtggcga ataagccacc atcaaaagga aagaacagga ctctggaggt
     2341 gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     2401 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     2461 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca tgcagccctc
     2521 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2581 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2641 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2701 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     2761 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2821 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2881 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2941 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     3001 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     3061 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     3121 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     3181 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     3241 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     3301 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     3361 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     3421 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     3481 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3541 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3601 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3661 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3721 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3781 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3841 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3901 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3961 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag gaagcacctc
     4021 cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc atcatcacca
     4081 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     4141 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     4201 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4261 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4321 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4381 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     4441 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4501 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4681 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4741 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4801 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4861 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4921 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4981 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5041 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5101 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5161 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5221 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5281 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5341 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6601 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6661 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6721 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6781 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6841 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6901 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6961 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     7021 ttcccagtca cgacgtt
//
