LOCUS       sequence-270418-        5888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector TMKL1 X013_pECIA14 should be used with bait vector TMKL1
            X013_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5888)
  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 5888)
  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_pECIA14
FEATURES             Location/Qualifiers
     source          1..5888
                     /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             2882..2905
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2921..2938
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2961..2978)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3011..3145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3035..3054)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3089..3108
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3289..3305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3289..3305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3302..3324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3313..3329
                     /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(3337..3343,3344..3361,3362..3367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3382..3403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3520..3537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3771..3790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4450..5241,5242..5310))
                     /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     5073..5092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5311..5415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5483..5501
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5539..5561)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5661..5680
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5874..5888,1..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 gggaccaagc ggcaatcccg gcggcgatct ggccccccag atgctgcgcg agctgcagga
     1261 gaccaatgcc gccctgcagg acgtgcgcga gctgctgcgc caccgcgtga aggagatcac
     1321 cttcctgaag aacaccgtga tggagtgcga cgcctgcggc atgcagccct ctagaggctc
     1381 cggctccatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     1441 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     1501 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     1561 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     1621 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     1681 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     1741 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     1801 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     1861 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     1921 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     1981 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     2041 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     2101 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     2161 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     2221 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     2281 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     2341 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     2401 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     2461 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     2521 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     2581 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     2641 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     2701 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     2761 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     2821 ccccgccggc accaccgacg ccgcgcaccc gggttctaga ggaagcacct ccggcgataa
     2881 cgattacaag gatgatgatg ataaggccag cggcaccggt catcatcacc atcaccattg
     2941 agtttaaacc cgctgatcag cctcgactgt gccttctaag gcctgagctc gctgatcagc
     3001 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3061 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3121 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3181 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     3241 tgattatgat cagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     3301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4561 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4741 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4921 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5401 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5461 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5521 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5581 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5641 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5701 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5761 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5821 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5881 acgacgtt
//
