LOCUS       sequence-442630-        6315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing HyperNova in mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6315)
  AUTHORS   Shidara H, Shirai T, Ozaki-Noma R, Jitsuki S, Nagai T, Takemoto K
  TITLE     Optical inactivation of intracellular molecules by fast-maturating 
            photosensitizing fluorescence protein, HyperNova.
  JOURNAL   Commun Biol. 2024 Aug 6;7(1):945. doi: 10.1038/s42003-024-06583-x.
  PUBMED    39107369
REFERENCE   2  (bases 1 to 6315)
  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: "Commun 
            Biol."; date: "2024-08-6"; volume: "7(1):945. doi"; pages: " 
            10.1038/s42003-024-06583-x"
COMMENT     Sequence Label: 2MLS-HyperNova/pcDNA3
FEATURES             Location/Qualifiers
     source          1..6315
                     /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             899..973
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             995..1066
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(1088..1090,1091..1096,1097..1807)
                     /codon_start=1
                     /product="monomeric variant of the photosensitizing red 
                     fluorescent protein KillerRed (Takemoto et al., 2013)"
                     /label=SuperNova
                     /translation="MGSEVGPALFQTDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGIIHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDGTCVVSRITVNCDGFLPDGPIMRDQLVDILPSETRILPYGPNAVRQTA
                     TIGFTTADGGKVMGHFDSKMTFNGSRAIEIPGPHFVTIITEQTRDPSDKRDHVCQREVA
                     YAHSGPRNTSAIGSDED"
     promoter        complement(1868..1886)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1869..1886)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1906..1923)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1912..2136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2182..2610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2269..2288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2479..2500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2619..2639)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2624..2953
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2804..2939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2866..2885
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3020..3814
                     /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(3074..3093)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3684..3703
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3988..4109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4025..4044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4079..4098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4158..4174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4158..4174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4171..4193)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4182..4198
                     /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(4206..4212,4213..4230,4231..4236))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4251..4272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4389..4406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4560..5148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4640..4659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5319..6110,6111..6179))
                     /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     5942..5961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6180..6284)
                     /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 gcttcaccat
      901 gtccgtcctg acgccgctgc tgctgcgggg cttgacaggc tcggcccggc ggctcccagt
      961 gccgcgcgcc aagatccatt cgttggggga tctgtccgtc ctgacgccgc tgctgctgcg
     1021 gggcttgaca ggctcggccc ggcggctccc agtgccgcgc gccaagatcc attcgttggg
     1081 gggatccatg ggttcggagg tcggccccgc cctgttccag accgacatga ccttcaaaat
     1141 cttcatcgac ggcgaggtta acggccagaa gttcaccatc gtggccgacg gcagcagcaa
     1201 gttcccccac ggcgacttca acgtgcacgc cgtgtgcgag accggcaagc tgcccatgag
     1261 ctggaagccc atctgccacc tgatccagta cggcgagccc ttcttcgccc gctaccccga
     1321 cggcatcatc catttcgccc aggagtgctt ccccgagggc ctgagcatcg accgcaccgt
     1381 gcgcttcgag aacgacggca ccatgaccag ccaccacacc tacgagctgg acggcacctg
     1441 cgtggtgagc cgcatcaccg tgaactgcga cggcttcctg cccgacggcc ccatcatgcg
     1501 cgaccagctg gtggacatcc tacccagcga gacccgcata ctcccctacg gccccaacgc
     1561 cgtgcgccag accgccacca tcggcttcac caccgccgac ggcggcaagg tgatgggcca
     1621 cttcgacagc aagatgacct tcaacggcag ccgcgccatc gagatccccg gcccacactt
     1681 cgtgaccatc atcaccgagc agacgaggga ccctagcgac aagcgcgacc acgtgtgcca
     1741 gcgcgaggtg gcctacgccc acagcgggcc ccgcaacacc agcgccatcg gtagcgacga
     1801 ggattaagaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg
     1861 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     1921 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1981 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2041 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2101 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2161 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2221 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2281 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2341 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2401 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2461 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2521 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2581 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     2641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2701 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2761 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2821 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2881 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2941 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3001 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3061 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3121 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3181 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3241 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3301 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3361 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3421 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3481 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3541 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3601 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3661 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3721 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3781 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3841 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     3901 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     3961 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4021 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4081 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4141 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4201 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4261 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4321 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6301 gtgccacctg acgtc
//
