LOCUS       sequence-274519-        7007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT1G29730 X066_pECIA14 should be used with bait vector 
            AT1G29730 X066_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7007)
  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 7007)
  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: AT1G29730 X066_pECIA14
FEATURES             Location/Qualifiers
     source          1..7007
                     /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             2303..2326
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4001..4024
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4040..4057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4080..4097)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4130..4264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4154..4173)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4208..4227
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4408..4424)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4408..4424)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4421..4443)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4432..4448
                     /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(4456..4462,4463..4480,4481..4486))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4501..4522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4639..4656)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4810..5398)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4890..4909)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5569..6360,6361..6429))
                     /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     6192..6211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6430..6534)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6602..6620
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6658..6680)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6780..6799
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6993..7007,1..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 tctccttctc tgcacccaga
      541 tgaagtggaa gcgctgaagg atatcactga aacattgggt gtgaagcact tgaacctaag
      601 tgaagatcca tgtctcacaa agactctagt gatatctcaa ggtgttctca aggaaggaca
      661 gaacagcaca atcagatgtg actgtcattt caataactac agcacttgtc atatcaaaca
      721 ctttgtcctc cagaaattca atcttccagg tagacttcct ccaatgctgt acaagtttcg
      781 gcatcttgaa tcgattgact tatacaataa ttacctttat ggctcaattc ctatggaatg
      841 ggcctcactg ccttacctca aatctatctc tgtctgtgca aaccgcttat caggagacat
      901 tcccaaagga ttggggaagt ttattaacct caccctttta gttcttgaag ccaatcagtt
      961 ttcgggaact attcctaagg aactagggaa cctagtgaat ctacaaggac taggactctc
     1021 ttccaaccaa cttgtcggag gcctccccaa gacattagca aaactaacaa agctaactaa
     1081 tctccattta agtgataatc gcctgaacgg atcgattcca gagtttattg ggaagttacc
     1141 aaaacttcaa agattagaac tctatgcgag tggccttaga ggacctattc cagactccat
     1201 ttttcatctg gagaatttga tcgacgtgag aatcagtgac acggttgcag gattgggaca
     1261 tgttcctcag ataactagca cgagcctgaa atatttggtt ttgagaaaca taaacttatc
     1321 tggaccaatt ccaaccagta tctgggatct gcccagttta atgactctgg atctttcctt
     1381 taataggctg actggagaaa taccagcata tgcaactgcc ccaaaataca catatttagc
     1441 tggaaacatg ttgtctggaa aggttgaaac aggagctttc cttactgcaa gcacaaatat
     1501 tgatctttcc tataataatt tcacgtggtc tccaatgtgc aaagagagga agaatataaa
     1561 cacatatgag agctcacact caaaaaaccg tctaaccaga cttcttccat gttccgccat
     1621 aaagcaatgc caaaattata gtagatcgct gcatataaac tgtgggggac ctgatgtaac
     1681 catcgaaaac tctcgaggaa ggtttctgta tgaaggtgat aactatggac ttaccggatc
     1741 agctacgaac tattatagga aaaattgggg atacagcaat actggtgact ttatggatga
     1801 tgcaataaca gaagatacat acacagtttc atcagaatct gcagtttcag caaaatatcc
     1861 agacctttac cagaatgctc ggcgctctcc cctcagtttg gcttactatg cattttgctt
     1921 cgaaaatgga agctacaatg tgaaactcca ttttgcagag attcagttct cagatgtgga
     1981 accatacact aaactagcga aacgggtttt taacatttac attcagggga agttgatttg
     2041 ggaggatttc agcattagag aggaggctaa tggaactcac aaagaagtta taagagaagt
     2101 aaacacgacc gtgactgata acactttaga gatacggctt tactgggcag ggaaaggcac
     2161 aatgatcatt cctcaaagag ggtactatgg ctctcttatc tctgcagtct cagtttgtcc
     2221 cagttcagaa tctgaatgcg gtggtatgaa gaagaaaatc tcaaagctta agggacctga
     2281 tctaagaaca ggttccttca gtctggaggt gctgttccag ggaccaagcg gcaatcccgg
     2341 cggcgatctg gccccccaga tgctgcgcga gctgcaggag accaatgccg ccctgcagga
     2401 cgtgcgcgag ctgctgcgcc accgcgtgaa ggagatcacc ttcctgaaga acaccgtgat
     2461 ggagtgcgac gcctgcggca tgcagccctc tagaggctcc ggctccatca tcccagttga
     2521 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     2581 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     2641 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     2701 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     2761 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     2821 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     2881 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     2941 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     3001 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     3061 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     3121 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     3181 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     3241 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     3301 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     3361 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     3421 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     3481 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     3541 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     3601 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     3661 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     3721 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     3781 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     3841 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     3901 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     3961 cgcgcacccg ggttctagag gaagcacctc cggcgataac gattacaagg atgatgatga
     4021 taaggccagc ggcaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     4081 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     4141 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4201 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4261 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     4321 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     4381 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4441 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4501 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4561 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4621 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4681 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4741 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4801 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4861 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4921 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4981 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5041 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5101 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5161 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5221 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5281 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5341 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5401 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5461 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5521 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5581 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5641 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5701 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5761 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5821 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5881 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5941 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6001 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6061 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6121 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6181 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6241 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6301 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6361 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6421 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6481 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6541 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     6601 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     6661 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6721 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6781 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6841 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6901 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6961 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
