LOCUS       sequence-334508-        7097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemo-optogenetic control of necroptotic cell death by coupling 
            LiPOP1 with NanoLuc-aided bioluminescence optogenetic stimulation 
            (NanoLOGS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7097)
  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 7097)
  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-NanoLuc
FEATURES             Location/Qualifiers
     source          1..7097
                     /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
                     EGRLSTGGMDELYK"
     primer_bind     complement(1078..1096)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1346..1365
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1659..2171
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    3825..3946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3862..3881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3916..3935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3953..4408)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4090..4111)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4302..4321
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4435..4539
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4541..4898
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4541..4732
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4564..4584)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4749..4884
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4933..5727
                     /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             4933..5727
                     /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(4987..5006)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5597..5616
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5915..5934)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5959..6006
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6335..6923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6824..6843
                     /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 gggccgcctc tccaccggcg
     1621 gcatggacga gctgtacaag ggtagcggac caccggtcat ggtcttcaca ctcgaagatt
     1681 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
     1741 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
     1801 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     1861 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
     1921 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     1981 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     2041 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     2101 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     2161 gcattctggc gccaccggtg cggccgcatt taattaaaat gaagatggac aaaaagacca
     2221 tcgtctggtt tcggagagat ttgagaatag aagataatcc cgcgctcgcc gccgcggccc
     2281 acgagggttc cgtcttcccc gttttcattt ggtgtcctga agaagaaggc cagttttatc
     2341 ccggaagggc ctctaggtgg tggatgaagc aaagtctggc ccatcttagc cagtcactga
     2401 aagcactggg cagtgatctt accctgatca agacacacaa taccatctct gccatcctcg
     2461 actgcatcag ggtgaccggc gcaacgaaag tcgtgtttaa ccacctgtac gatccagtta
     2521 gtctggtgcg cgaccacact gtgaaggaga agctggtgga acgggggatc agtgtgcaga
     2581 gctacaacgg ggaccttctg tacgagccat gggagatcta ttgcgagaaa gggaaaccgt
     2641 tcacctcctt caacagttac tggaagaaat gtttggatat gtcaatagag tccgttatgt
     2701 tgccccctcc ctggagactg atgccgatta ctgctgctgc agaggccatc tgggcctgct
     2761 ccatcgagga actcggtctg gaaaatgaag cagaaaagcc aagcaatgca cttctcacta
     2821 gagcctggag ccccggctgg tctaatgccg acaagctgct taacgagttc atcgaaaaac
     2881 aactgattga ctacgcgaag aactccaaga aagtggtagg taactcaact agcttgctct
     2941 ctccatatct ccattttggc gagatttctg tccgccatgt atttcagtgc gctcggatga
     3001 aacagattat ctgggctcgc gataaaaaca gcgaaggcga agaaagcgcc gatctgttcc
     3061 tgcgagggat cggacttcgg gaatactccc ggtatatatg tttcaacttt ccattcacac
     3121 acgagcagag tctgttgtcc cacctcaggt tcttcccctg ggacgccgat gtcgacaaat
     3181 tcaaggcatg gagacaggga aggacagggt acccactcgt ggatgctggc atgagagagc
     3241 tctgggctac aggctggatg cacaaccgca tccgggtaat cgtgtcctca tttgctgtca
     3301 agtttctgct cctgccttgg aaatggggaa tgaagtactt ttgggatacc cttctcgacg
     3361 ccgacttgga gtgtgacatt ctgggatggc aatatattag cgggtcaatt cctgacggcc
     3421 atgagttgga caggttggac aatccggcct tgcagggagc taagtatgat cccgaaggag
     3481 agtatattcg acagtggctc cccgagctgg cccgacttcc tacggagtgg attcaccatc
     3541 cttgggacgc accactgaca gtgctcaagg caagcggggt ggagctgggc accaattacg
     3601 ctaagcctat agttgatata gatacagcac gcgagctgct ggctaaagcg atctctcgca
     3661 ctcgggaggc gcagattatg atcggtgctg cctagggatc caccggatct agataactga
     3721 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3781 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3841 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3901 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     3961 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4021 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4081 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4141 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4201 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4261 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4321 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4381 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4501 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4561 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4621 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4681 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4741 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4801 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4861 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4921 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4981 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5041 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5101 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5161 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5221 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5281 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5341 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5401 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5461 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5521 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5581 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5641 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5701 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5761 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5821 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5881 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5941 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6001 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6061 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6121 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6181 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6241 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6301 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6421 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6481 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6541 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6601 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6661 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6721 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6781 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6841 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6901 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6961 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7021 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     7081 ccatatatgg agttccg
//
