LOCUS       sequence-298847-        6287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector SKM2 X057_pECIA2 should be used with prey vector SKM2 
            X057_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6287)
  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 6287)
  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: SKM2 X057_pECIA2
FEATURES             Location/Qualifiers
     source          1..6287
                     /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             2540..2563
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3269..3310
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3320..3337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3360..3377)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3410..3544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3434..3453)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3488..3507
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3688..3704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3688..3704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3701..3723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3712..3728
                     /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(3736..3742,3743..3760,3761..3766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3781..3802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3919..3936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4090..4678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4170..4189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4849..5640,5641..5709))
                     /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     5472..5491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5710..5814)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5882..5900
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5938..5960)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6060..6079
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6273..6287,1..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 tccatagacg aacagggtct
      541 agctctcctc tcatggaaat ctcagttgaa catctccgga gacgctttat cttcttggaa
      601 agcctccgaa tcgaatcctt gtcaatgggt gggaatcaaa tgcaacgaaa gaggtcaagt
      661 ctcggagata caactccaag tcatggattt tcaaggtccg ttaccggcga cgaatctccg
      721 acagataaag tctctgactt tgctcagctt aacctccgtg aatctcactg gttcaatccc
      781 taaggagctt ggagacttgt cggagctaga agttcttgat ttagctgata attctctctc
      841 tggtgaaatc cctgtggata tcttcaagct caagaaacta aagattctgt ctctcaacac
      901 aaataatctc gaaggtgtga ttccatcgga gttagggaat ctcgttaacc tcatcgagct
      961 cactctcttc gacaataaac tcgccggaga gattcctaga accatcggag aactcaagaa
     1021 tctagaaatc ttccgtgcag gtggtaacaa gaatctcaga ggagagcttc cttgggagat
     1081 tggtaactgc gagagtcttg tgactttagg tctcgccgaa acaagtctct ccggtagact
     1141 tccggcgtcg atcggaaacc tgaaaaaggt tcagacaata gctctttaca cgtcactctt
     1201 gtctggtcca atccctgatg agattggaaa ctgcacagag cttcaaaacc tctacttgta
     1261 ccaaaactca atctctggat cgattcctgt atcaatggga cgtctcaaga agcttcaaag
     1321 tcttctctta tggcaaaaca atctcgtcgg aaagatccct accgagctag gcacctgtcc
     1381 ggagcttttc ctcgtcgatt tatcggaaaa tctcctcacc ggaaacatcc caagaagctt
     1441 cggaaacctt ccaaatctcc aagagcttca gctaagtgtt aaccagctct ccggtacaat
     1501 ccctgaagaa ctcgcgaatt gcacgaagtt gacgcattta gagatcgata acaaccaaat
     1561 ctccggcgag attccgccgt tgatcgggaa attaacaagc ttgacgatgt tctttgcgtg
     1621 gcagaatcag ttaaccggaa taattccaga gtctctttct caatgtcaag agcttcaagc
     1681 gattgatctc tcctacaaca atctctctgg ttcgatacct aatggaatct ttgagattcg
     1741 aaacctcacg aagcttcttc ttctctctaa ctacttatct ggattcatac caccggatat
     1801 tggaaactgc acgaatcttt accggttaag actcaacggt aacagactcg cggggaatat
     1861 tccggcggag attgggaatc tgaaaaatct caacttcatc gatataagtg agaatcgtct
     1921 catcggaaac attcctccgg agatctccgg ttgtacaagt ctcgaattcg tcgatctaca
     1981 ttcaaacggt ttaaccggtg gtttacccgg tacacttccg aagagcttac aattcattga
     2041 tttatccgat aattcattaa ccggttctct accaaccgga atcggatcgt taaccgaact
     2101 cacgaagctc aatctcgcga agaatcgttt ctccggcgaa atccctcgag agatatcttc
     2161 ctgtcggagt cttcagcttc tcaacctcgg cgacaatggc ttcaccggag aaatccctaa
     2221 cgagttagga cggatcccgt ctcttgcgat ctctctaaat ctcagctgca atcacttcac
     2281 cggagagatc ccgtcgagat tctctagcct cactaatctc ggaaccctcg atgtctctca
     2341 caacaaactc gccggaaacc taaacgtctt agctgatcta cagaacctcg tctcactcaa
     2401 catctccttc aacgaattct caggcgagtt accaaatact ctgtttttca ggaaacttcc
     2461 actctctgtt cttgaatcaa acaaagggct atttatttcg acccgacccg agaatgggat
     2521 ccaaacccga caccgttcgc tggaggtgct gttccaggga ccagggagcg acaaaactca
     2581 cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc
     2641 cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt
     2701 ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt
     2761 gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag
     2821 cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc
     2881 caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg
     2941 agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag
     3001 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     3061 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     3121 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     3181 atgctccgtg atgcatgagg gtctgcacaa ccactacacg cagaagagcc tctccctgtc
     3241 tccgggtaaa tctagagggc ccttcgaagg taagcctatc cctaaccctc tcctcggtct
     3301 cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     3361 ctcgactgtg ccttctaagg cctgagctcg ctgatcagcc tcgatcgagg atccagacat
     3421 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3481 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3541 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3601 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     3661 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5821 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5881 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     5941 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     6001 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     6061 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     6121 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     6181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     6241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgtt
//
