LOCUS       sequence-270153-        6677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector MEE39 N5_pECIA14 should be used with bait vector MEE39 
            N5_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6677)
  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 6677)
  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: MEE39 N5_pECIA14
FEATURES             Location/Qualifiers
     source          1..6677
                     /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             1973..1996
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3671..3694
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3710..3727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3750..3767)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3800..3934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3824..3843)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3878..3897
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4078..4094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4078..4094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4091..4113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4102..4118
                     /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(4126..4132,4133..4150,4151..4156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4171..4192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4309..4326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4480..5068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4560..4579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5239..6030,6031..6099))
                     /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     5862..5881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6100..6204)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6272..6290
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6328..6350)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6450..6469
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6663..6677,1..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 caagaaggtt ttatcagttt
      541 agattgtggc ttacctctaa atgagccacc ttacatagaa tcggaaaccg gaatacagtt
      601 ctcatcagac gagaatttca tccaaagtgg aaagaccggt agaatcccga aaaatcttga
      661 atcggaaaat ttaaaacagt atgcaaccct acgatacttt ccagatggaa tacgaaactg
      721 ctatgatcta agggtggaag aagggaggaa ctaccttatt agagctacat ttttctatgg
      781 aaattttgac gggcttaatg ttagccctga attcgatatg catatcggtc ctaataagtg
      841 gacaacaata gatttgcaaa tagtgccaga tggtacagtg aaggagatca ttcacattcc
      901 aagatcaaac tctttgcaaa tttgtcttgt taagacaggg gcaactatac caatgatatc
      961 agctttggaa ttacgacctt tggcgaatga tacttacatc gctaaatctg gttccttgaa
     1021 atattatttt cgcatgtatc ttagcaatgc caccgttctt cttcggtacc cgaaagatgt
     1081 ttatgatcgt tcgtgggttc catatattca acccgaatgg aatcaaattt caacaacttc
     1141 aaatgtgtcc aataaaaatc actatgatcc accacaagtt gctcttaaga tggccgcaac
     1201 gcctacaaat cttgatgcag cactgacaat ggtttggcgt ttggagaatc ctgatgatca
     1261 aatctacttg tacatgcact tctcggagat ccaagtccta aaagccaatg acactagaga
     1321 attcgacatt atattgaatg gagaaacaat taacactaga ggtgttactc ctaaatatct
     1381 tgaaataatg acctggctta caacaaatcc taggcaatgc aacggaggta tctgccgaat
     1441 gcagttaaca aaaacccaaa aatcgacact tccacctcta cttaatgctt ttgaagttta
     1501 ctcagttctc cagcttccac aatctcagac aaatgaaatt gaagtggttg ctatcaaaaa
     1561 cattagaact acttatggac tgagtagaat cagttggcaa ggagatccat gcgtaccgaa
     1621 acaattcttg tgggatggat tgaattgtaa catcacagat atatctgcac caccaagaat
     1681 aatttcttta aatttatcat cgagtggctt gagtgggact atagtgtcta actttcagaa
     1741 tctggcgcat ctagaaagtt tggatttgtc aaataacagt ttgagtggaa tagtgcccga
     1801 gtttctagcc actatgaaat cattgttggt cataaacttg agcgggaata aacttagtgg
     1861 tgcaattcct caagctcttc gcgatagaga aagggagggg ctaaagttaa atgttcttgg
     1921 aaacaaagag ctttgtctat ctagtacatg catagataag ccaaagaaaa aactggaggt
     1981 gctgttccag ggaccaagcg gcaatcccgg cggcgatctg gccccccaga tgctgcgcga
     2041 gctgcaggag accaatgccg ccctgcagga cgtgcgcgag ctgctgcgcc accgcgtgaa
     2101 ggagatcacc ttcctgaaga acaccgtgat ggagtgcgac gcctgcggca tgcagccctc
     2161 tagaggctcc ggctccatca tcccagttga ggaggagaac ccggacttct ggaaccgcga
     2221 ggcagccgag gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa
     2281 cctcatcatc ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct
     2341 aaaagggcag aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc
     2401 atatgtggct ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac
     2461 agccacggcc tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc
     2521 cgcccgcttt aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg
     2581 ggccaagaaa gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc
     2641 gccagccggc acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc
     2701 tgcctcggcc cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga
     2761 cattgacgtg atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc
     2821 tgagtaccca gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca
     2881 ggaatggctg gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca
     2941 ggcttccctg gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa
     3001 atacgagatc caccgagact ccacactgga cccctccctg atggagatga cagaggctgc
     3061 cctgcgcctg ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat
     3121 cgaccatggt catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga
     3181 cgacgccatt gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac
     3241 tgccgaccac tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt
     3301 cgggctggcc cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa
     3361 cggtccaggc tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg
     3421 gagccccgag tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga
     3481 ggacgtggcg gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca
     3541 gaccttcata gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga
     3601 cctggcgccc cccgccggca ccaccgacgc cgcgcacccg ggttctagag gaagcacctc
     3661 cggcgataac gattacaagg atgatgatga taaggccagc ggcaccggtc atcatcacca
     3721 tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg cctgagctcg
     3781 ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt ttggacaaac
     3841 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3901 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3961 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4021 tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt aatcatggtc
     4081 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4141 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4201 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4261 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4321 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4381 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4441 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4501 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4561 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4621 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4681 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4741 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4801 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4861 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4921 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4981 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5041 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5101 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5161 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5221 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5281 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5341 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5401 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5461 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5521 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5581 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5641 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5701 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5761 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5821 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5881 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5941 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6001 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6061 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6121 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6181 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     6241 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     6301 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     6361 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6421 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6481 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     6541 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6601 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6661 ttcccagtca cgacgtt
//
