LOCUS       sequence-270343-        6827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT2G27060 B10_pECIA14 should be used with bait vector 
            AT2G27060 B10_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6827)
  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 6827)
  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: AT2G27060 B10_pECIA14
FEATURES             Location/Qualifiers
     source          1..6827
                     /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             2123..2146
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3821..3844
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3860..3877
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3900..3917)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3950..4084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3974..3993)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4028..4047
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4228..4244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4228..4244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4241..4263)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4252..4268
                     /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(4276..4282,4283..4300,4301..4306))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4321..4342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4459..4476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4630..5218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4710..4729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5389..6180,6181..6249))
                     /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     6012..6031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6250..6354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6422..6440
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6478..6500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6600..6619
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6813..6827,1..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 tctgactttg aagctctttt
      541 ggagcttaag aaaggttttc agggtgaccc ttctcgaaaa gttcttactt catgggatgc
      601 taaagcttta tcctctgata ggtgtccgtt aaactggtac ggagttactt gtagtagcgg
      661 aggtgttact tctattgatt tgaatggttt cggtctgctt gggagtttca gttttccagt
      721 cattgttggt ctgagaatgc ttcagaattt gtcgattgcg aataatcagt ttagtggaac
      781 tctctctaac attggttcgt taacgtctct taagtacttg gatgtttcgg gtaacttgtt
      841 ccatggcgca ttaccttctg gaattgagaa tttgaggaac cttgagtttg tgaatctctc
      901 tggtaacaac aacttgggag gagtgattcc ttctggtttt gggagtcttg ccaagttgaa
      961 gtaccttgat ttgcaaggca acagtttctc tggggaagtg atgagtctgt tttcacagct
     1021 aattagtgtg gagtatgtag atataagcag gaacaacttc tcgggttcgc ttgatctggg
     1081 tctagcaaag tcaagctttg tttcctcgat tcgtcatttg aacgtaagtg ggaattcttt
     1141 ggtaggggag ctatttgctc atgatggtat cccctttttc gatagcttag aggtgtttga
     1201 tgctagcagc aatcaattgt ctggttctgt tccagtcttc agctttgttg tctctctcaa
     1261 gattcttcgg ttacaagaca atcagttgtc ggcttcttta ccaccgggcc ttctgcaaga
     1321 gagttcaacc atcttaactg accttgatct tagcctcaat caacttgaag gtcctatcgg
     1381 aagtataacc tcttcaacac tagagaagct caatttgtct tcaaatagac tatcgggatc
     1441 attacctttg aaagttggac attgtgccat tattgatctg agtaacaata agatatcagg
     1501 agaattgtca aggatacaga attggggaga ttctgtagaa attattcggt tgagttcaaa
     1561 ctcactcact ggaacactac ctggccaaac ttctcaattc ctgaggctga cttcactcaa
     1621 ggctgctaac aactcgttgc agggtgtact accatttatt ttaggaacgt atcctgagct
     1681 gaaagagatt gatctcagtc acaaccagct tagtggcgtc ataccgagta atctgttcat
     1741 atctgcaaag ttgacggaac tcaacttgtc aaacaataat ttctctggtt ctcttcctct
     1801 tcaagatgca agtactgtgg gaaaccttag cttgaccaat attggtctgt ctcataactc
     1861 gttaggtgga gttctctcag aggaactaac tcgcttccac aatctgattt ctcttgacct
     1921 ttcttataac aactttgaag ggaatatccc tgatggcctt ccagacagcc taaagatgtt
     1981 cactgtctct gccaataatc tctctggaaa tgtgccggag aatcttagac ggtttccaga
     2041 ttcagcgttt catccaggga acgctttact gaatgtcccc atttctctac caaaagataa
     2101 aacagatata accttacgga agctggaggt gctgttccag ggaccaagcg gcaatcccgg
     2161 cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg ccctgcagga
     2221 cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat
     2281 ggagtgcgac gcctgcggca tgcagccctc tagaggctcc ggctccatca tcccagttga
     2341 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     2401 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     2461 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     2521 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     2581 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     2641 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     2701 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     2761 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     2821 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     2881 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     2941 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     3001 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     3061 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     3121 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     3181 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     3241 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     3301 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     3361 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     3421 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     3481 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     3541 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     3601 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     3661 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     3721 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     3781 cgcgcacccg ggttctagag gaagcacctc cggcgataac gattacaagg atgatgatga
     3841 taaggccagc ggcaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     3901 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     3961 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4021 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4081 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4141 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     4201 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4261 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4321 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4381 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4441 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4501 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4561 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5041 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5341 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5401 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5461 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5521 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5581 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5641 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5701 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5761 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5821 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5881 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5941 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6001 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6061 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6121 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6181 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6241 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6361 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6421 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6481 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6541 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6601 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6661 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6721 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6781 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
