LOCUS       sequence-270166-        6686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT4G29450 O1_pECIA14 should be used with bait vector 
            AT4G29450 O1_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6686)
  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 6686)
  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: AT4G29450 O1_pECIA14
FEATURES             Location/Qualifiers
     source          1..6686
                     /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"
     RBS             550..558
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1982..2005
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3680..3703
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3719..3736
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3759..3776)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3809..3943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3833..3852)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3887..3906
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4087..4103)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4087..4103)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4100..4122)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4111..4127
                     /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(4135..4141,4142..4159,4160..4165))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4180..4201
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4318..4335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4489..5077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4569..4588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5248..6039,6040..6108))
                     /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     5871..5890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6109..6213)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6281..6299
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6337..6359)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6459..6478
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6672..6686,1..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 tcaggttaca tcagcatcga
      541 ttgtggcata cctccttacg atacccctga agacactatg actaacatta attacgtctc
      601 cgatgaagct ttcataacca ccggcgtaaa cttcaaggtc tctgaagaat acggctatcc
      661 aaaaaacccg gtattgctgt caactcttgc tgaagtaaga gctttccctc aaggaaacag
      721 aaactgttac accttaaaac tttcacaagg aaaagatcat ctttatctca ttagagcttc
      781 ctttatgtat ggaaactacg atggtaaaaa agctttgcct gagtttgatc tctatgtaaa
      841 cgtgaacttc tggtcgacag tgaaattcaa aaatgcttct gatcaagtca ccaaagagat
      901 tttaagcttt gcagagtcgg atacaatata tgtttgcctt gtgaataaag gcaaaggaac
      961 tccttttatt tcagggttag agctccgacc agtgaacagc tcaatctacg gtactgaatt
     1021 tggaagaaat gtctctttgg ttctctatcg aagatgggac attggatatt taaacggaac
     1081 aggacggtac caagacgata gattcgatcg tatttggtct ccatactcct caaatatctc
     1141 ctggaactcg attataacat cgggctatat cgatgttttt cagaacggtt attgtccacc
     1201 tgatgaagtt attaagacag ctgctgctcc tgaaaacgtt gatgatccgt tggaattgtt
     1261 ctggacatca gacgatccaa atgttcggtt ctacgcatat ctttacttcg cggaactcga
     1321 aacactagag aagaatgaga ctagaaagat caagattttg tggaatggtt cccctgtttc
     1381 tgaaacttct ttcgaacctt cttcaaaata ttcaacgacg ttttctaatc caagagcttt
     1441 cactggtaaa gatcactgga tttctattca gaagactgta gattcaacgc ttccaccgat
     1501 tctcaatgct atagagattt ttaccgcaca atctctagat gaattttcca ccacaattga
     1561 agacattcat gctatagaaa gcataaaagc aacttataaa gtgaacaagg tttggagtgg
     1621 tgatccttgc tccccaagat tattcccttg ggaaggtgtt ggatgcagcg acaacaataa
     1681 taatcatcaa atcaagtccc tgaatcttag ctcaagcgga ttactaggac caatagtctt
     1741 ggcatttaga aatctctctc ttcttgagtc attggattta tccaacaatg acctgcaaca
     1801 aaatgtgcct gagtttttgg ctgatctaaa acatttgaaa gtcctgaatt tgaaaggaaa
     1861 caattttact gggtttatcc caaaatccct tatgaaaaag ttaaaggccg gtttacttac
     1921 gctcagtgcg gatgagcaaa acctttgtaa ctcatgtcaa gaaaagaaga agaagaagag
     1981 cctggaggtg ctgttccagg gaccaagcgg caatcccggc ggcgatctgg ccccccagat
     2041 gctgcgcgag ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc tgctgcgcca
     2101 ccgcgtgaag gagatcacct tcctgaagaa caccgtgatg gagtgcgacg cctgcggcat
     2161 gcagccctct agaggctccg gctccatcat cccagttgag gaggagaacc cggacttctg
     2221 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     2281 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     2341 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     2401 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     2461 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     2521 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     2581 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     2641 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     2701 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     2761 caacatggac attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac
     2821 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     2881 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga
     2941 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     3001 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     3061 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     3121 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     3181 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     3241 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     3301 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     3361 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     3421 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     3481 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     3541 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     3601 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gttctagagg
     3661 aagcacctcc ggcgataacg attacaagga tgatgatgat aaggccagcg gcaccggtca
     3721 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     3781 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     3841 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3901 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3961 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4021 ctacaaatgt ggtatggctg attatgatca gtcgacctgc aggcatgcaa gcttggcgta
     4081 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4141 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4201 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4261 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4321 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4381 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4441 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4501 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4561 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4621 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4681 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4741 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4801 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4861 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4921 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4981 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5041 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5101 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5161 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5221 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5281 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5341 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5401 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5461 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5521 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5581 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5641 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5701 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5761 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5821 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5881 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5941 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6001 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6061 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6121 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6181 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6241 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6301 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6361 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6421 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6481 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6541 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6601 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6661 gccagggttt tcccagtcac gacgtt
//
