LOCUS       sequence-322540-        6259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for caspase-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6259)
  AUTHORS   Lu X, Wen Y, Zhang S, Zhang W, Chen Y, Shen Y, Lemieux MJ, Campbell 
            RE
  TITLE     Photocleavable proteins that undergo fast and efficient 
            dissociation.
  JOURNAL   Chem Sci. 2021 May 31;12(28):9658-9672. doi: 10.1039/d1sc01059j. 
            eCollection 2021 Jul 21.
  PUBMED    34349937
REFERENCE   2  (bases 1 to 6259)
  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: 
            "10.1039/d1sc01059j"; journalName: "Chem Sci"; date: "2021-05-31- 
            31"; volume: "12"; issue: "28"; pages: "9658-9672"
COMMENT     Sequence Label: pcDNA-NES-DEVD-mCardinal-NLS
FEATURES             Location/Qualifiers
     source          1..6259
                     /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             997..1725
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQR
                     IKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCDM
                     ALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     CDS             1735..1755
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1759..1779
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1783..1803
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1850..1867)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1856..2080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2126..2554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2213..2232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2423..2444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2563..2583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2568..2897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2748..2883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2810..2829
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2964..3758
                     /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(3018..3037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3628..3647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3932..4053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3969..3988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4023..4042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4102..4118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4102..4118)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4115..4137)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4126..4142
                     /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(4150..4156,4157..4174,4175..4180))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4195..4216
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4333..4350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4504..5092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4584..4603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5263..6054,6055..6123))
                     /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     5886..5905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6124..6228)
                     /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 gccaccatga acctggtgga cctgcagaag aagctggagg agctggagct ggacgagcag
      961 cagggatccg cctccggcga tgaggtggat ggagccgtga gcaagggcga ggagctgatc
     1021 aaggagaaca tgcacatgaa gctgtacatg gaaggcaccg tgaacaacca ccacttcaag
     1081 tgcaccaccg aaggggaggg caagccctac gagggcaccc agacccagag gattaaggtg
     1141 gtggagggag gccccctgcc gttcgcattc gacatcctgg ccacctgctt tatgtacggg
     1201 agcaagacct tcatcaacca cacccagggc atccccgatt tctttaagca gtccttccct
     1261 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct taccgttacc
     1321 caggacacca gcctccagga cggctgcttg atctacaacg tcaagctcag aggggtgaac
     1381 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc caccaccgag
     1441 accctgtacc ccgctgacgg cggcctggaa ggcagatgcg acatggccct gaagctcgtg
     1501 ggcgggggcc acctgcactg caacctgaag accacataca gatccaagaa acccgctaag
     1561 aacctcaaga tgcccggcgt ctactttgtg gaccgcagac tggaaagaat caaggaggcc
     1621 gacaatgaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct
     1681 agcaaactgg ggcacaaact taatggcatg gacgagctgt acaagggctc cggaccaaaa
     1741 aaaaaacgaa aagttgaccc aaaaaagaag cgcaaagtag accccaagaa aaaacgcaag
     1801 gtaggtacct aactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1861 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1921 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1981 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2041 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2101 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2161 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2221 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2281 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2341 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2401 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2461 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2521 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2581 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2641 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2701 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2761 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2821 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2881 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     2941 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3001 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3061 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3121 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3181 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3241 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3301 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3361 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3421 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3481 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3541 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3601 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3661 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3721 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3781 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3841 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3901 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3961 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4021 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4081 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4141 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4201 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4261 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4321 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4381 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4441 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4501 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4561 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4621 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4681 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4741 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4801 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4861 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4921 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4981 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5041 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5101 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5161 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5221 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5281 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5341 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5401 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5461 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5521 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5581 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5641 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5701 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5761 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5821 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5881 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5941 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6001 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6061 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6121 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6181 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6241 aaaagtgcca cctgacgtc
//
