LOCUS       sequence-296827-        4931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector TMKL1 X013_pECIA2 should be used with prey vector TMKL1 
            X013_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4931)
  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 4931)
  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: TMKL1 X013_pECIA2
FEATURES             Location/Qualifiers
     source          1..4931
                     /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             1184..1207
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1913..1954
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1964..1981
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2004..2021)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2054..2188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2078..2097)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2132..2151
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2332..2348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2332..2348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2345..2367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2356..2372
                     /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(2380..2386,2387..2404,2405..2410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2425..2446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2563..2580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2734..3322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2814..2833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3493..4284,4285..4353))
                     /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     4116..4135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4354..4458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4526..4544
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4582..4604)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4704..4723
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4917..4931,1..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 tctctctctg gttcttctga
      541 tgtgaagctt cttttaggaa aaatcaagtc ttcactacaa ggaaacagtg agagcttact
      601 gttgtcttct tggaactcct ctgttcctgt ttgtcaatgg agaggtgtaa aatgggtatt
      661 ttcaaatggg tctcctcttc aatgtagtga cctctcttca ccacaatgga ctaatacctc
      721 tctgttcaac gactcttctc ttcaccttct ctctcttcag cttccttctg ctaatctcac
      781 tggttcactc cctagagaga ttggtgagtt ctctatgctt caaagtgtgt tcctcaacat
      841 caattcatta agtgggtcaa tccctcttga gcttggttac acttcttctc tctctgatgt
      901 tgatttgagt ggtaatgcct tagctggggt tttgcctcca tcgatttgga acctctgtga
      961 taagcttgtc tctttcaaga ttcatggtaa taacttgtct ggggttttgc ctgagcctgc
     1021 tttgccaaat tcgacttgtg gtaatctcca agttcttgat ttgggtggta ataagttctc
     1081 agattttggt gagtcaaagt ttggagcaga atctttcgaa gggaacagtc ctagcctttg
     1141 tggtttgcct ttgaagcctt gtctaggctc ctctaggtta agtctggagg tgctgttcca
     1201 gggaccaggg agcgacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg
     1261 gggaccgtca gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac
     1321 ccctgaggtc acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa
     1381 ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta
     1441 caacagcacg taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg
     1501 caaggagtac aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat
     1561 ctccaaagcc aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga
     1621 ggagatgacc aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga
     1681 catcgccgtg gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc
     1741 cgtgctggac tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag
     1801 gtggcagcag gggaacgtct tctcatgctc cgtgatgcat gagggtctgc acaaccacta
     1861 cacgcagaag agcctctccc tgtctccggg taaatctaga gggcccttcg aaggtaagcc
     1921 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     1981 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     2041 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     2101 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2161 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2221 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     2281 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     2341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2461 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2521 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2581 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4501 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4561 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4621 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4681 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4741 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     4801 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4861 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     4921 gtcacgacgt t
//
