LOCUS       sequence-296607-        5732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector FRK1/SIRK M10_pECIA2 should be used with prey vector 
            FRK1/SIRK M10_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5732)
  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 5732)
  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: FRK1/SIRK M10_pECIA2
FEATURES             Location/Qualifiers
     source          1..5732
                     /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             1985..2008
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2714..2755
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2765..2782
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2805..2822)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2855..2989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2879..2898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2933..2952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3133..3149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3133..3149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3146..3168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3157..3173
                     /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(3181..3187,3188..3205,3206..3211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3226..3247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3364..3381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3535..4123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3615..3634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4294..5085,5086..5154))
                     /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     4917..4936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5155..5259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5327..5345
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5383..5405)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5505..5524
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5718..5732,1..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 tctggtttca taagtataga
      541 ttgcgggata ccggatgatt ctagctacaa cgatgagact acaggtataa agtatgtttc
      601 ggattcggcg tttgttgatt caggaacaac aaagagaatt gcagctcagt ttcaatcaag
      661 tggttttgat agacacttgt tgaacgtgag aagtttccct caaagcaaga gaagctgtta
      721 cgatgttccg acgccgagag gcaaaggttt taagtatcta atcagaactc gtttcatgta
      781 cgggaactat gatgatcttg gaagagtacc cgagttcgat ctctatctag gagtaaactt
      841 ttgggactct gttaaactcg acgatgcaac aactatactc aacaaagaga taatcaccat
      901 tccactttta gacaatgttc aagtgtgtgt tgttgataag aacgcaggaa ctccattttt
      961 gtctgtcttg gagatacggc tcttgttgaa cactacttat gagactcctt acgatgcact
     1021 taccctcctt cggagattag attacagcaa aacgggaaaa cttccatcga ggtacaaaga
     1081 tgacatatat gaccgtatat ggacaccgcg tatagtgagt tcagaataca agatattaaa
     1141 tacatctctc actgtcgatc aatttcttaa caatggctac caaccggctt ctactgtcat
     1201 gagcactgca gaaacagcgc gaaacgagag cctctaccta acgcttagtt tcagaccgcc
     1261 cgaccctaac gcaaagtttt atgtatacat gcactttgct gaaattgaag tactgaaaag
     1321 caaccagacg agagaattca gcatctggct aaatgaggat gtaatctctc cttcgtttaa
     1381 gcttcggtac ttgcttaccg acacattcgt tacaccggat cccgtgagcg gaattaccat
     1441 taacttctct cttcttcaac cacctggtga atttgtactt ccaccgatca ttaacgccct
     1501 tgaggtctat caagtcaatg agttccttca gatcccaact catccacagg atgttgatgc
     1561 catgaggaag attaaagcca cgtatagagt gaagaagaac tggcaaggag atccttgtgt
     1621 tcccgtagat tattcttggg aaggtattga ctgtatccaa agtgataaca ctactaatcc
     1681 tagagtcgtt tcactaaata tatcttttag tgaattacga ggccagatag atccagcctt
     1741 ctccaacctt acatctataa gaaaattaga tttatccggt aatactttaa caggagaaat
     1801 acctgctttc ctcgctaact taccaaactt gaccgaatta aacgttgaag gaaacaagtt
     1861 aacgggcata gttccacaaa gattgcatga aagatcaaag aatggatctc tttccttaag
     1921 atttggtaga aatccggacc tttgtctctc tgattcctgt tcaaacacaa agaagaagaa
     1981 caagctggag gtgctgttcc agggaccagg gagcgacaaa actcacacat gcccaccgtg
     2041 cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa aacccaagga
     2101 caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg tgagccacga
     2161 agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata atgccaagac
     2221 aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc tcaccgtcct
     2281 gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca aagccctccc
     2341 agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac cacaggtgta
     2401 caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga cctgcctggt
     2461 caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc agccggagaa
     2521 caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc tctacagcaa
     2581 gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct ccgtgatgca
     2641 tgagggtctg cacaaccact acacgcagaa gagcctctcc ctgtctccgg gtaaatctag
     2701 agggcccttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     2761 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2821 taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga
     2881 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2941 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3001 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3061 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     3121 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3181 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3241 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3301 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3361 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3421 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3481 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3721 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3901 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3961 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4021 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4081 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4141 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4201 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4261 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4321 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4381 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4441 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4501 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4561 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4621 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4681 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4741 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4801 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4861 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4921 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4981 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5041 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5101 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5161 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5221 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5281 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     5341 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5401 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5461 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     5521 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5581 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5641 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5701 ggtaacgcca gggttttccc agtcacgacg tt
//
