LOCUS       sequence-107782-        7922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses infrared fluorescent executioner caspase reporter 
            (iCasper) plus human heme oxygenase 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7922)
  AUTHORS   To TL, Piggott BJ, Makhijani K, Yu D, Jan YN, Shu X
  TITLE     Rationally designed fluorogenic protease reporter visualizes 
            spatiotemporal dynamics of apoptosis in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3338-43. doi: 
            10.1073/pnas.1502857112. Epub 2015 Mar 2.
  PUBMED    25733847
REFERENCE   2  (bases 1 to 7922)
  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: 
            "10.1073/pnas.1502857112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-17- 17"; volume: "112"; issue: "11"; pages: "3338-43"
COMMENT     Sequence Label: pcDNA3.1 iCasper T2A HO1
FEATURES             Location/Qualifiers
     source          1..7922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..1564
                     /codon_start=1
                     /product="fragment of superfolder GFP consisting of the 
                     first 10 beta-strands"
                     /label=GFP(1-10)
                     /note="associates with GFP11, the 11th beta-strand, to 
                     reconstitute fluorescent GFP (Cabantous et al., 2005; 
                     Kamiyama et al., 2016)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     CDS             2507..2554
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             2564..2617
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(3516..3533)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3522..3746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3792..4220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3879..3898)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4089..4110
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4229..4249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4234..4563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4414..4549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4476..4495
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4630..5424
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4684..4703)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5294..5313
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5598..5719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5635..5654)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5689..5708
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5768..5784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5768..5784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5781..5803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5792..5808
                     /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(5816..5822,5823..5840,5841..5846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5861..5882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5999..6016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6170..6755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6250..6269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6926..7717,7718..7786))
                     /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     7549..7568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7787..7891)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgtccaa aggagaagaa ctgtttaccg gtgttgtgcc
      961 aattttggtt gaactcgatg gtgatgtcaa cggacataag ttctcagtga gaggcgaagg
     1021 agaaggtgac gccaccattg gaaaattgac tcttaaattc atctgtacta ctggtaaact
     1081 tcctgtacca tggccgactc tcgtaacaac gcttacgtac ggagttcagt gcttttcgag
     1141 atacccagac catatgaaaa gacatgactt ttttaagtcg gctatgcctg aaggttacgt
     1201 gcaagaaaga acaatttcgt tcaaagatga tggaaaatat aaaactagag cagttgttaa
     1261 atttgaagga gatactttgg ttaaccgcat tgaactgaaa ggaacagatt ttaaagaaga
     1321 tggtaatatt cttggacaca aactcgaata caattttaat agtcataacg tatacatcac
     1381 tgctgataag caaaagaacg gaattaaagc gaatttcaca gtacgccata atgtagaaga
     1441 tggcagtgtt caacttgccg accattacca acaaaacacc cctattggag acggtccggt
     1501 acttcttcct gataatcact acctctcaac acaaacagtc ctgagcaaag atccaaatga
     1561 aaaaggaggc agcggatcca ccactaacat tgcgccggct ctggacggtg cgtttcatcg
     1621 tatcacttct tcatcctccc tgatgggcct gtgtgacgaa accgcgacta ttatccgtga
     1681 gattactggc tacgaccgtg tgatggtagt acgtttcgat gaagagggta atgccgaaat
     1741 tctgtccgaa cgtcgtcgtg cggacctgga agcgttcctg ggtaaccgct acccggcgtc
     1801 tactattccg cagatcgctc gtcgcctgta cgaacataac cgtgttcgcc tgctggtaga
     1861 tgtgaactat actccggttc cgctacagcc gcgcatcagc ccgctgaacg gtcgtgatct
     1921 ggatatgtcc ctgtcttgcc tgcgctctat gtccccggtc caccagaaat acatgcagga
     1981 catgggcgtt ggcgcgaccc tggtttgctc tctgatggtg tctggtcgtt tgtggggtct
     2041 gatcgcttgc caccactacg aaccgcgctt cgttccgttc cacattcgcg ctgctggcga
     2101 agcgctggcg gaaacttgtg cgaaccgcat cgcgacgctg gagagcggtg atgaagttga
     2161 tggtgaattc tgtgaacgcg agcagatcca cctggcgggc tccattcagc cgcacggtat
     2221 cctgctggct gtgaaagagc cagacaacgt ggtgatccag gcttctatca acgctgcgga
     2281 gttcctgaac accaactctg ttgttggccg tccgctgcgt gacctgggcg gcgatctgcc
     2341 tttgcagatc ctgccgcacc tgaacggccc gctgcacctg gctccggtga ccctgcgttg
     2401 taccgtgggt tctccgccgc gccgtgtgga ctgtaccatt caccgtccgt ctaacggcgg
     2461 cctgatcgta gaactggaac cagcaaccaa gctcgaggga ggcagccggg accacatggt
     2521 gctgcacgag tacgtgaacg ccgccggcat cacagggcgc gccgagggca gaggaagtct
     2581 gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg gaaagaccac agccagactc
     2641 tatgccacag gatctgagcg aagccctcaa ggaggccacc aaggaagtgc acacccaggc
     2701 cgagaacgct gagttcatga gaaacttcca gaaaggacag gttactcgcg acggattcaa
     2761 gctggtgatg gctagcctct atcacatcta cgtggccctg gaggaggaaa ttgagagaaa
     2821 caaggaatcc cccgtgttcg ctccagttta tttcccagag gagctgcatc ggaaggccgc
     2881 cctggaacag gatctggctt tttggtatgg cccccgctgg caggaggtca ttccttatac
     2941 ccccgctatg caacgctacg ttaagagact ccacgaggtg ggccgcacag aacctgagct
     3001 gctcgtggcc catgcctaca cccgctacct gggcgacctg tcagggggtc aagttctgaa
     3061 aaagatagcc caaaaggccc tggacctgcc ttcctctggc gaggggctgg ccttctttac
     3121 tttccctaac attgcctcag ccacaaagtt caagcagctc taccggtcca ggatgaacag
     3181 cctggaaatg accccagccg tgcggcagcg ggtcatagaa gaagccaaga ctgcctttct
     3241 gctgaatatc cagctgtttg aggaactcca ggaactgctg acccatgaca ctaaagacca
     3301 gtccccctcc agggccccag gactcagaca gagggcctca aacaaggttc aggactcagc
     3361 ccccgtggaa acacccaggg gaaaaccccc tctcaataca agatctcaag cacccctgct
     3421 gagatgggtc ctcaccctga gcttcctcgt cgctaccgtt gcagtggggc tctatgctat
     3481 gtgatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3541 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3601 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3661 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3721 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3781 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3841 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3901 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3961 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4021 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4081 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4141 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4201 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4261 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4381 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4441 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4501 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4561 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4621 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4681 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4741 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4801 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4861 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4921 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4981 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5041 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5101 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5161 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5221 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5281 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5341 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5401 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5461 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5521 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5581 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5641 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5701 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5761 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5821 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5881 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5941 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6001 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6061 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6721 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6781 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6841 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6901 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6961 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7021 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7081 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7141 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7201 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7261 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7321 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7381 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7441 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7501 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7561 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7621 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7681 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7741 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7801 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7861 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7921 tc
//
