LOCUS       sequence-270260-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G65240 I5_pECIA14 should be used with bait vector 
            AT5G65240 I5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  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 5816)
  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: AT5G65240 I5_pECIA14
FEATURES             Location/Qualifiers
     source          1..5816
                     /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             1112..1135
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2810..2833
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2849..2866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2889..2906)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2939..3073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2963..2982)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3017..3036
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3217..3233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3217..3233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3230..3252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3241..3257
                     /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(3265..3271,3272..3289,3290..3295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3310..3331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3448..3465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3619..4207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3699..3718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4378..5169,5170..5238))
                     /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     5001..5020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5239..5343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5411..5429
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5467..5489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5589..5608
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5802..5816,1..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 tcaccagatg ctcaagggga
      541 tgcattattt gcgttgagga gctcgttacg tgcatctcct gaacagctta gtgattggaa
      601 ccagaatcaa gtcgatcctt gtacttggtc tcaagttatt tgtgatgaca agaaacatgt
      661 tacttctgta accttgtctt acatgaactt ctcctcggga acactgtctt caggaatagg
      721 aatcttgaca actctcaaga ctcttacatt gaagggaaat ggaataatgg gtggaatacc
      781 agaatccatt ggaaatctgt ctagcttgac cagcttagat ttggaggata atcacttaac
      841 tgatcgcatt ccatccactc tcggtaatct caagaatcta cagttcttga ccttgagtag
      901 gaataacctt aatggttcta tcccggattc acttacaggt ctatcaaaac tgataaatat
      961 tctgctcgac tcaaataatc tcagtggtga gattcctcag agtttattca aaatcccaaa
     1021 atacaatttc acagcaaaca acttgagctg tggtggcact ttcccgcaac cttgtgtaac
     1081 cgagtccagt ccttcaggtg attcaagcag tctggaggtg ctgttccagg gaccaagcgg
     1141 caatcccggc ggcgatctgg ccccccagat gctgcgcgag ctgcaggaga ccaatgccgc
     1201 cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag gagatcacct tcctgaagaa
     1261 caccgtgatg gagtgcgacg cctgcggcat gcagccctct agaggctccg gctccatcat
     1321 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     1381 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     1441 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     1501 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     1561 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     1621 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     1681 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     1741 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     1801 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     1861 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     1921 aggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     1981 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     2041 gggtgcccgg tatgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     2101 gacccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     2161 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     2221 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     2281 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     2341 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     2401 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     2461 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     2521 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     2581 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     2641 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     2701 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     2761 caccgacgcc gcgcacccgg gttctagagg aagcacctcc ggcgataacg attacaagga
     2821 tgatgatgat aaggccagcg gcaccggtca tcatcaccat caccattgag tttaaacccg
     2881 ctgatcagcc tcgactgtgc cttctaaggc ctgagctcgc tgatcagcct cgatcgagga
     2941 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3001 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3061 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3121 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     3181 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3241 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3301 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3361 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3421 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3481 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5341 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5401 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5461 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5521 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5581 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5641 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5701 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5761 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
