LOCUS       sequence-343558-        7391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  localizes KillerRed to mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7391)
  AUTHORS   Kidwell CU, Casalini JR, Pradeep S, Scherer SD, Greiner D, Bayik D, 
            Watson DC, Olson GS, Lathia JD, Johnson JS, Rutter J, Welm AL, 
            Zangle TA, Roh-Johnson M
  TITLE     Transferred mitochondria accumulate reactive oxygen species, 
            promoting proliferation.
  JOURNAL   eLife. 2023 Mar 6;12:e85494. doi: 10.7554/eLife.85494.
  PUBMED    36876914
REFERENCE   2  (bases 1 to 7391)
  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: "eLife."; 
            date: "2023-03-6"; volume: "12:e85494. doi"; pages: " 
            10.7554/eLife.85494"
COMMENT     Sequence Label: pLKO.1 3xHA-KillerRed-OMP25
FEATURES             Location/Qualifiers
     source          1..7391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             572..598
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             611..637
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             650..676
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(689..691,692..697,698..1405)
                     /codon_start=1
                     /product="red fluorescent protein KillerRed, a 
                     genetically-encoded photosensitizer (Bulina et al., 2006)"
                     /label=KillerRed
                     /note="mammalian codon-optimized"
                     /translation="MGSEGGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDDTCVVSRITVNCDGFQPDGPIMRDQLVDILPNETHMFPHGPNAVRQLA
                     FIGFTTADGGLMMGHFDSKMTFNGSRAIEIPGPHFVTIITKQMRDTSDKRDHVCQREVA
                     YAHSVPRITSAIGSDED"
     LTR             1597..1830
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1902..2023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1939..1958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1993..2012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2063..2198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2218..2237)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2219..2237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2247..2264)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2247..2263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2256..2278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2405..2860
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2492..2511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2702..2723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2886..2990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2991..3059,3060..3851)
                     /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     complement(3209..3228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4022..4610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4511..4530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4764..4781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4898..4919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4934..4939,4940..4957,4958..4964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4988
                     /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)."
     primer_bind     4977..4999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4996..5012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4996..5012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5031..5051
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5033..5051
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5079..5305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5306..5486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5533..5658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6151..6384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6569..6613
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        6911..7159
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6911..6931
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7082..7101
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    7222..7339
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(7388..7391,1..507)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccgacgt cttaattaac ctgcagggtc
      541 gacacgcgtc gtacgaccgg agcttaccat gtatccctat gacgtgcctg attacgccgg
      601 cacaggatcc tacccctatg atgtgcctga ctacgctggc agcgccggat acccttatga
      661 tgtgcctgat tatgctggag ggagcggcat gggttcagag ggcggccccg ccctgttcca
      721 gagcgacatg accttcaaaa tcttcatcga cggcgaggtg aacggccaga agttcaccat
      781 cgtggccgac ggcagcagca agttccccca cggcgacttc aacgtgcacg ccgtgtgcga
      841 gaccggcaag ctgcccatga gctggaagcc catctgccac ctgatccagt acggcgagcc
      901 cttcttcgcc cgctaccccg acggcatcag ccatttcgcc caggagtgct tccccgaggg
      961 cctgagcatc gaccgcaccg tgcgcttcga gaacgacggc accatgacca gccaccacac
     1021 ctacgagctg gacgacacct gcgtggtgag ccgcatcacc gtgaactgcg acggcttcca
     1081 gcccgacggc cccatcatgc gcgaccagct ggtggacatc ctgcccaacg agacccacat
     1141 gttcccccac ggccccaacg ccgtgcgcca gctggccttc atcggcttca ccaccgccga
     1201 cggcggcctg atgatgggcc acttcgacag caagatgacc ttcaacggca gccgcgccat
     1261 cgagatcccc ggcccacact tcgtgaccat catcaccaag cagatgaggg acaccagcga
     1321 caagcgcgac cacgtgtgcc agcgcgaggt ggcctacgcc cacagcgtgc cccgcatcac
     1381 cagcgccatc ggtagcgacg aggatccgcg gcatcgaggc gacggagagc ctagtggagt
     1441 tcctgtagct gtggtgctgc tgccagtgtt tgcccttacc ctggtagcag tttgggcctt
     1501 cgtgagatac cgaaagcagc tctgaggtac ctttaagacc aatgacttac aaggcagctg
     1561 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     1621 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     1681 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     1741 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1801 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     1861 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     1921 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1981 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     2041 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2101 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2161 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     2221 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     2281 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2341 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2401 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2461 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2521 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2581 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2641 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2701 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2761 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2821 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2881 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2941 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3001 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3061 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3121 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3181 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3241 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3301 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3361 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3421 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3481 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3541 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3601 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3661 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3721 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3781 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3841 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4801 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4861 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4921 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4981 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     5041 ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt
     5101 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
     5161 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
     5221 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
     5281 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
     5341 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     5401 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     5461 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     5521 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     5581 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     5641 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     5701 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     5761 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     5821 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     5881 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     5941 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     6001 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     6061 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     6121 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     6181 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     6241 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     6301 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     6361 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     6421 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     6481 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     6541 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     6601 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     6661 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     6721 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     6781 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     6841 cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc gagcggccgc
     6901 ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     6961 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     7021 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     7081 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     7141 gtggaaagga cgaaacaccg gtccgcaggt atgcacgcgt gaattctcga cctcgagaca
     7201 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     7261 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     7321 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac tttggccgcg
     7381 gctcgagggg g
//
