LOCUS       sequence-238699-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector BIR4 C2_pECIA14 should be used with bait vector BIR4 
            C2_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  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 5810)
  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: BIR4 C2_pECIA14
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        468..894
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     912..965
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1551..1574
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3249..3272
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3288..3305
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3328..3345)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3378..3512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3402..3421)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3456..3475
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3656..3672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3656..3672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3669..3691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3680..3696
                     /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(3704..3710,3711..3728,3729..3734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3749..3770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3887..3904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4058..4646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4138..4157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4817..5608,5609..5677))
                     /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     5440..5459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5678..5782)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcgttgcagg
      481 acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat cctctggttc
      541 cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat acatatgtgg
      601 tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt tgcatcccat
      661 acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa agcttctgca
      721 cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa tcgaacgaaa
      781 gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct ggccaatgtg
      841 catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta aaggggggat
      901 ccgatctcaa tatgaagtta tgcatattac tggccgtcgt ggcctttgtt ggcctctcgc
      961 tcgggtctca cgcagaagac gatgtactct gtctcaaagg ctttaaatca tcactcaaag
     1021 atccttcaaa tcaactcaac acatggtctt tccctaattc atcatcgtcg atttgcaaac
     1081 tcaccggagt ttcttgctgg aacgctaaag agaatcgaat tctctctctt cagcttcaat
     1141 caatgcaact ctctggtcaa atccctgaat ctcttaaact atgtcggagt ttacaatctt
     1201 tagatctttc ctttaatgat ttctccggtt tgattccttc acaaatatgt tcttggcttc
     1261 cttatcttgt tactttagat ctttccggta acaaactctc cggttcgata ccgtctcaga
     1321 tcgttgactg taaattctta aacagcttag ctctaaacca gaacaagtta accggttcga
     1381 ttccgtctga attaacccgg ttaaaccgtc tccaacgact ttctctagct gataacgatc
     1441 tctctggttc gattccctcc gagctatctc attacggaga agatggtttt cgcggtaatg
     1501 gcggactttg tggtaagccg ttatccaact gcggttcgtt taacggtaaa ctggaggtgc
     1561 tgttccaggg accaagcggc aatcccggcg gcgatctggc cccccagatg ctgcgcgagc
     1621 tgcaggagac caatgccgcc ctgcaggacg tgcgcgagct gctgcgccac cgcgtgaagg
     1681 agatcacctt cctgaagaac accgtgatgg agtgcgacgc ctgcggcatg cagccctcta
     1741 gaggctccgg ctccatcatc ccagttgagg aggagaaccc ggacttctgg aaccgcgagg
     1801 cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc acagacagcc gccaagaacc
     1861 tcatcatctt cctgggcgat gggatggggg tgtctacggt gacagctgcc aggatcctaa
     1921 aagggcagaa gaaggacaaa ctggggcctg agatacccct ggccatggac cgcttcccat
     1981 atgtggctct gtccaagaca tacaatgtag acaaacatgt gccagacagt ggagccacag
     2041 ccacggccta cctgtgcggg gtcaagggca acttccagac cattggcttg agtgcagccg
     2101 cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt catctccgtg atgaatcggg
     2161 ccaagaaagc agggaagtca gtgggagtgg taaccaccac acgagtgcag cacgcctcgc
     2221 cagccggcac ctacgcccac acggtgaacc gcaactggta ctcggacgcc gacgtgcctg
     2281 cctcggcccg ccaggagggg tgccaggaca tcgctacgca gctcatctcc aacatggaca
     2341 ttgacgtgat cctaggtgga ggccgaaagt acatgtttcg catgggaacc ccagaccctg
     2401 agtacccaga tgactacagc caaggtggga ccaggctgga cgggaagaat ctggtgcagg
     2461 aatggctggc gaagcgccag ggtgcccggt atgtgtggaa ccgcactgag ctcatgcagg
     2521 cttccctgga cccgtctgtg acccatctca tgggtctctt tgagcctgga gacatgaaat
     2581 acgagatcca ccgagactcc acactggacc cctccctgat ggagatgaca gaggctgccc
     2641 tgcgcctgct gagcaggaac ccccgcggct tcttcctctt cgtggagggt ggtcgcatcg
     2701 accatggtca tcatgaaagc agggcttacc gggcactgac tgagacgatc atgttcgacg
     2761 acgccattga gagggcgggc cagctcacca gcgaggagga cacgctgagc ctcgtcactg
     2821 ccgaccactc ccacgtcttc tccttcggag gctaccccct gcgagggagc tccatcttcg
     2881 ggctggcccc tggcaaggcc cgggacagga aggcctacac ggtcctccta tacggaaacg
     2941 gtccaggcta tgtgctcaag gacggcgccc ggccggatgt taccgagagc gagagcggga
     3001 gccccgagta tcggcagcag tcagcagtgc ccctggacga agagacccac gcaggcgagg
     3061 acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg caggagcaga
     3121 ccttcatagc gcacgtcatg gccttcgccg cctgcctgga gccctacacc gcctgcgacc
     3181 tggcgccccc cgccggcacc accgacgccg cgcacccggg ttctagagga agcacctccg
     3241 gcgataacga ttacaaggat gatgatgata aggccagcgg caccggtcat catcaccatc
     3301 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct
     3361 gatcagcctc gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca
     3421 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3481 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3541 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3601 gtatggctga ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat
     3661 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3721 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3781 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3841 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4681 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4741 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4801 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4861 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4921 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4981 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5041 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5101 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5161 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5221 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5281 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5341 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5401 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5461 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5521 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5581 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5641 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5701 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5761 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
