LOCUS       sequence-296632-        5720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bait vector MEE39 N5_pECIA2 should be used with prey vector MEE39 
            N5_pECIA14..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5720)
  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 5720)
  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_pECIA2
FEATURES             Location/Qualifiers
     source          1..5720
                     /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             2702..2743
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2753..2770
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2793..2810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2843..2977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2867..2886)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2921..2940
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3121..3137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3121..3137)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3134..3156)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3145..3161
                     /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(3169..3175,3176..3193,3194..3199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3214..3235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3352..3369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3523..4111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3603..3622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4282..5073,5074..5142))
                     /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     4905..4924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5143..5247)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5315..5333
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5371..5393)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5493..5512
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5706..5720,1..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 ggaccaggga gcgacaaaac tcacacatgc ccaccgtgcc cagcacctga
     2041 actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat
     2101 ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt
     2161 caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga
     2221 ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg
     2281 gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga
     2341 gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc
     2401 atcccgggag gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta
     2461 tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac
     2521 cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga
     2581 caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg agggtctgca
     2641 caaccactac acgcagaaga gcctctccct gtctccgggt aaatctagag ggcccttcga
     2701 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
     2761 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta aggcctgagc
     2821 tcgctgatca gcctcgatcg aggatccaga catgataaga tacattgatg agtttggaca
     2881 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2941 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3001 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3061 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
     3121 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3181 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3241 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3301 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3361 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3961 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4261 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4321 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4381 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4441 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4501 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4561 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4621 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4681 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4741 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4801 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4861 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4921 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4981 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5221 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     5281 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5341 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5401 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5461 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5521 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5581 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5641 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     5701 gttttcccag tcacgacgtt
//
