LOCUS       sequence-334504-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A single-component optogenetic tool for photoswitchable necroptosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  AUTHORS   He L, Huang Z, Huang K, Chen R, Nguyen NT, Wang R, Cai X, Huang Z, 
            Siwko S, Walker JR, Han G, Zhou Y, Jing J
  TITLE     Optogenetic Control of Non-Apoptotic Cell Death.
  JOURNAL   Adv Sci (Weinh). 2021 May 6;8(13):2100424. doi: 
            10.1002/advs.202100424. eCollection 2021 Jul.
  PUBMED    34540558
REFERENCE   2  (bases 1 to 6575)
  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"; journalName: "Adv Sci 
            (Weinh)."; date: "2021-05-6"; volume: "8(13):2100424. doi"; pages: "
            10.1002/advs.202100424. eCollection 2021 Jul"
COMMENT     Sequence Label: LiPOP1
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             933..1640
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1078..1096)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1346..1365
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3303..3424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3340..3359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3394..3413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3431..3886)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3568..3589)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3780..3799
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3913..4017
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4019..4376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4019..4210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4042..4062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4227..4362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4411..5205
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4411..5205
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4465..4484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5075..5094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5393..5412)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5437..5484
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5813..6401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6302..6321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      541 aaaatttgaa gcatattatc acccttggcc aggtcatcgc cgcagcgtgt gaagagatga
      601 aatactgcaa gaaacagtgc cggcgcctgg gccaccgcgt cctcggcctg atcaagcctc
      661 tggagatgct ccaggaccaa ggaaagagga gcgtgccctc tgagaagtta accacagcca
      721 tgaaccgctt caaggctgcc ctggaggagg ctaatgggga gatagaaaag ttcagcaata
      781 gatccaatat ctgcaggttt ctaacagcaa gccaggacaa aatactcttc aaggacgtga
      841 acaggaagct gagtgatgtc tggaaggagc tctcgctgtt acttcaggtt gagcaacgca
      901 tgcctgtttc aggaagcggt gctagcggtg gaatggtgag caagggcgag gaggataaca
      961 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     1021 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     1081 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     1141 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     1201 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     1261 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     1321 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1381 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     1441 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     1501 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     1561 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     1621 gcatggacga gctgtacaag ggtagcggac caccggtgcg gccgcattta attaaaatga
     1681 agatggacaa aaagaccatc gtctggtttc ggagagattt gagaatagaa gataatcccg
     1741 cgctcgccgc cgcggcccac gagggttccg tcttccccgt tttcatttgg tgtcctgaag
     1801 aagaaggcca gttttatccc ggaagggcct ctaggtggtg gatgaagcaa agtctggccc
     1861 atcttagcca gtcactgaaa gcactgggca gtgatcttac cctgatcaag acacacaata
     1921 ccatctctgc catcctcgac tgcatcaggg tgaccggcgc aacgaaagtc gtgtttaacc
     1981 acctgtacga tccagttagt ctggtgcgcg accacactgt gaaggagaag ctggtggaac
     2041 gggggatcag tgtgcagagc tacaacgggg accttctgta cgagccatgg gagatctatt
     2101 gcgagaaagg gaaaccgttc acctccttca acagttactg gaagaaatgt ttggatatgt
     2161 caatagagtc cgttatgttg ccccctccct ggagactgat gccgattact gctgctgcag
     2221 aggccatctg ggcctgctcc atcgaggaac tcggtctgga aaatgaagca gaaaagccaa
     2281 gcaatgcact tctcactaga gcctggagcc ccggctggtc taatgccgac aagctgctta
     2341 acgagttcat cgaaaaacaa ctgattgact acgcgaagaa ctccaagaaa gtggtaggta
     2401 actcaactag cttgctctct ccatatctcc attttggcga gatttctgtc cgccatgtat
     2461 ttcagtgcgc tcggatgaaa cagattatct gggctcgcga taaaaacagc gaaggcgaag
     2521 aaagcgccga tctgttcctg cgagggatcg gacttcggga atactcccgg tatatatgtt
     2581 tcaactttcc attcacacac gagcagagtc tgttgtccca cctcaggttc ttcccctggg
     2641 acgccgatgt cgacaaattc aaggcatgga gacagggaag gacagggtac ccactcgtgg
     2701 atgctggcat gagagagctc tgggctacag gctggatgca caaccgcatc cgggtaatcg
     2761 tgtcctcatt tgctgtcaag tttctgctcc tgccttggaa atggggaatg aagtactttt
     2821 gggataccct tctcgacgcc gacttggagt gtgacattct gggatggcaa tatattagcg
     2881 ggtcaattcc tgacggccat gagttggaca ggttggacaa tccggccttg cagggagcta
     2941 agtatgatcc cgaaggagag tatattcgac agtggctccc cgagctggcc cgacttccta
     3001 cggagtggat tcaccatcct tgggacgcac cactgacagt gctcaaggca agcggggtgg
     3061 agctgggcac caattacgct aagcctatag ttgatataga tacagcacgc gagctgctgg
     3121 ctaaagcgat ctctcgcact cgggaggcgc agattatgat cggtgctgcc tagggatcca
     3181 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3241 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3301 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3361 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3421 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3481 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3541 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3601 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3661 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3721 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3781 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3841 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3901 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3961 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4021 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4081 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4141 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4201 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4381 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4441 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4501 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4561 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     4621 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4681 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4741 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4801 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4861 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4921 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4981 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5041 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5101 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5161 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5221 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5281 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5341 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     5401 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     5461 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     5521 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     5581 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     5641 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5701 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5761 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5821 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5881 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5941 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6001 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6061 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6121 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6181 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6241 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6301 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6361 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6421 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6481 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     6541 gggtcattag ttcatagccc atatatggag ttccg
//
