LOCUS       sequence-260104-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondria matrix localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4739)
  TITLE     Okada Lab plasmids
REFERENCE   2  (bases 1 to 4739)
  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"
COMMENT     Sequence Label: pN1-COX8-mScarletI
FEATURES             Location/Qualifiers
     source          1..4739
                     /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             537..611
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             645..1340
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(778..796)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1046..1065
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1467..1588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1504..1523)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1558..1577
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1595..2050)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1732..1753)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1944..1963
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2077..2181
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2183..2540
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2183..2374
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2206..2226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2391..2526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2575..3369
                     /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             2575..3369
                     /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(2629..2648)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3239..3258
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3557..3576)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3601..3648
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3977..4565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4466..4485
                     /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 gctagcatgt
      541 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc
      601 cgcgcgccaa gatccattcg ttgggggatc caccggtcgc caccatggtg agcaagggcg
      661 aggcagtgat caaggagttc atgcggttca aggtgcacat ggagggctcc atgaacggcc
      721 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      781 agctgaaggt gaccaagggt ggccccctgc ccttctcctg ggacatcctg tcccctcagt
      841 tcatgtacgg ctccagggcc ttcatcaagc accccgccga catccccgac tactataagc
      901 agtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg
      961 tgaccgtgac ccaggacacc tccctggagg acggcaccct gatctacaag gtgaagctcc
     1021 gcggcaccaa cttccctcct gacggccccg taatgcagaa gaagacaatg ggctgggaag
     1081 cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt aagatggccc
     1141 tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa gaccacctac aaggccaaga
     1201 agcccgtgca gatgcccggc gcctacaacg tcgaccgcaa gttggacatc acctcccaca
     1261 acgaggacta caccgtggtg gaacagtacg aacgctccga gggccgccac tccaccggcg
     1321 gcatggacga gctgtacaag ttaaagcggc cgcgactcta gatcataatc agccatacca
     1381 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1441 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1501 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1561 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     1621 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1681 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     1741 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1801 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     1861 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     1921 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     1981 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2041 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2161 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2221 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2281 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2341 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2521 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2581 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2641 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2701 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     2761 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2821 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2881 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2941 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3001 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3061 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3121 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3181 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3241 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3301 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3361 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3421 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3481 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3541 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3601 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     3661 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3721 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3781 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     3841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4621 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     4681 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
