LOCUS       sequence-274755-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector BIR3 B11_pECIA14 should be used with bait vector BIR3 
            B11_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: BIR3 B11_pECIA14
FEATURES             Location/Qualifiers
     source          1..5810
                     /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             1106..1129
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2804..2827
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2843..2860
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2883..2900)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2933..3067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2957..2976)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3011..3030
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3211..3227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3211..3227)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3224..3246)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3235..3251
                     /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(3259..3265,3266..3283,3284..3289))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3304..3325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3442..3459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3613..4201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3693..3712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4372..5163,5164..5232))
                     /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     4995..5014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5233..5337)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5405..5423
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5461..5483)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5583..5602
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5796..5810,1..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 tcagctgaag acgacgttct
      541 ctgtctccaa ggtttgaaaa actcactcat cgatccttct tctagactca gttcctggtc
      601 tttcccaaac tcttcagctt cttcgatttg taagctcact ggtgtttcct gctggaacga
      661 gaaagagaat cggattattt ctcttcagct tcaatcgatg cagctcgccg gtgagattcc
      721 tgagtctctt aagctttgtc gtagtttaca atctctggat ctctctggaa acgatctctc
      781 tggttcgatt ccttctcaaa tctgttcgtg gcttccttat ctcgttacgt tagatctgtc
      841 tgggaataag cttggtggtt cgattccgac tcagatcgtt gagtgtaagt tcttaaacgc
      901 tttgattctg agtgataata agctttctgg ttcgattcct tctcagttga gtcgattaga
      961 tcgccttcga cgtctttctc tagcagggaa tgatctctcc ggtacgattc cgtccgagct
     1021 ggctcggttc ggaggagacg attttagcgg gaataacgga ctttgtggga agcctttatc
     1081 gcgatgtgga gcgttgaatg ggagactgga ggtgctgttc cagggaccaa gcggcaatcc
     1141 cggcggcgat ctggcccccc agatgctgcg cgagctgcag gagaccaatg ccgccctgca
     1201 ggacgtgcgc gagctgctgc gccaccgcgt gaaggagatc accttcctga agaacaccgt
     1261 gatggagtgc gacgcctgcg gcatgcagcc ctctagaggc tccggctcca tcatcccagt
     1321 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     1381 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     1441 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     1501 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     1561 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     1621 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     1681 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     1741 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     1801 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     1861 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     1921 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     1981 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     2041 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     2101 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     2161 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     2221 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     2281 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     2341 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     2401 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     2461 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     2521 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     2581 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     2641 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     2701 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     2761 cgccgcgcac ccgggttcta gaggaagcac ctccggcgat aacgattaca aggatgatga
     2821 tgataaggcc agcggcaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     2881 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     2941 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3001 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3061 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     3121 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     3181 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3241 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3301 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3361 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4021 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5341 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5401 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5461 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5521 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5581 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5641 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5701 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5761 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
