LOCUS       sequence-72833-e        4088 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses split fluorescence protein to measure caspase-2 
            dimerization in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4088)
  AUTHORS   Bouchier-Hayes L, Oberst A, McStay GP, Connell S, Tait SW, Dillon 
            CP, Flanagan JM, Beere HM, Green DR
  TITLE     Characterization of cytoplasmic caspase-2 activation by induced 
            proximity.
  JOURNAL   Mol Cell. 2009 Sep 24;35(6):830-40. doi: 
            10.1016/j.molcel.2009.07.023.
  PUBMED    19782032
REFERENCE   2  (bases 1 to 4088)
  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: 
            "10.1016/j.molcel.2009.07"; journalName: "Mol Cell"; date: 
            "2009-09-24- 24"; volume: "35"; issue: "6"; pages: "830-40"
COMMENT     Sequence Label: pBiFC-flag-hCaspase 2 FL (C303S)-VN173
FEATURES             Location/Qualifiers
     source          1..4088
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..30
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             133..384
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     321..342
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    399..533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(423..442)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     477..496
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(644..660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(644..660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(657..679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    668..684
                     /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(692..698,699..716,717..722))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    737..758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(875..892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1046..1634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1126..1145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1805..2596,2597..2665))
                     /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     2428..2447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2666..2770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2838..2856
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2894..2916)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3016..3035
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3229..3251
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3243..3260
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3244..3260
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3287..3590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3591..3794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3745..3765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3791..3815
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3932..4028
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 atgtacccat acgatgttcc agattacgct cttatggcca tggaggcccg aattcggtcg
       61 accgagatct ctcgaggtac ccgtccggcg tgcaaaatcc cgaacgacct gaaacagaaa
      121 gtcatgaacc acgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
      181 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      241 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
      301 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      361 ctcggcatgg acgagctgta caagtaagcg gccgcgggga tccagacatg ataagataca
      421 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
      481 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
      541 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttcggatc
      601 ctctagagtc gatctgcagg catgctagct tggcgtaatc atggtcatag ctgtttcctg
      661 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      721 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      781 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      841 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      901 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      961 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1561 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1621 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1681 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1741 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1801 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1861 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1921 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1981 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2041 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2101 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2161 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2221 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2281 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2341 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2401 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2461 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2521 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2581 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2641 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     2821 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2881 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3001 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     3061 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     3121 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     3181 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     3241 gttgtaaaac gacggccagt gagttcgagc ttgcatgcct gcaggtcgtt acataactta
     3301 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     3361 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     3421 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     3481 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     3541 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     3601 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     3661 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     3721 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     3781 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     3841 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagag gatccggtac
     3901 tagaggaact gaaaaaccag aaagttaact ggtaagttta gtctttttgt cttttatttc
     3961 aggtcccgga tccggtggtg gtgcaaatca aagaactgct cctcagtgga tgttgccttt
     4021 acttctaggc ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg
     4081 ggcccacc
//
