LOCUS       sequence-260106-        5025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondrial intermembrane space localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5025)
  TITLE     Okada Lab plasmids
REFERENCE   2  (bases 1 to 5025)
  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-OPA1(123Nt)-mScarletI
FEATURES             Location/Qualifiers
     source          1..5025
                     /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"
     regulatory      925..934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             931..1626
                     /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(1064..1082)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1332..1351
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1753..1874
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1790..1809)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1844..1863
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1881..2336)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2018..2039)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2230..2249
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2363..2467
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2469..2826
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2469..2660
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2492..2512)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2677..2812
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2861..3655
                     /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             2861..3655
                     /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(2915..2934)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3525..3544
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3843..3862)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3887..3934
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4263..4851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4752..4771
                     /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 gctagccacc
      541 atgtggcgac tacgtcgggc cgctgtggcc tgtgaggtct gccagtcttt agtgaaacac
      601 agctctggaa taaaaggaag tttaccacta caaaaactac atctggtttc acgaagcatt
      661 tatcattcac atcatcctac cttaaagctt caacgacccc aattaaggac atcctttcag
      721 cagttctctt ctctgacaaa ccttccttta cgtaaactga aattctctcc aattaaatat
      781 ggctaccagc ctcgcaggaa tttttggcca gcaagattag ctacgagact cttaaaactt
      841 cgctatctca tactaggatc ggctgttggg ggtggctaca cagccaaaaa gacttttgat
      901 cagtggaaag gggatccacc ggtcgccacc atggtgagca agggcgaggc agtgatcaag
      961 gagttcatgc ggttcaaggt gcacatggag ggctccatga acggccacga gttcgagatc
     1021 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1081 aagggtggcc ccctgccctt ctcctgggac atcctgtccc ctcagttcat gtacggctcc
     1141 agggccttca tcaagcaccc cgccgacatc cccgactact ataagcagtc cttccccgag
     1201 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgccgtgac cgtgacccag
     1261 gacacctccc tggaggacgg caccctgatc tacaaggtga agctccgcgg caccaacttc
     1321 cctcctgacg gccccgtaat gcagaagaag acaatgggct gggaagcgtc caccgagcgg
     1381 ttgtaccccg aggacggcgt gctgaagggc gacattaaga tggccctgcg cctgaaggac
     1441 ggcggccgct acctggcgga cttcaagacc acctacaagg ccaagaagcc cgtgcagatg
     1501 cccggcgcct acaacgtcga ccgcaagttg gacatcacct cccacaacga ggactacacc
     1561 gtggtggaac agtacgaacg ctccgagggc cgccactcca ccggcggcat ggacgagctg
     1621 tacaagttaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     1681 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     1741 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1801 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1861 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1921 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     1981 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2041 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2101 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2161 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2221 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2281 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     2341 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2401 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2461 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     2521 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2581 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2641 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2701 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2761 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2821 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     2881 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2941 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3001 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3061 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3121 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3181 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3241 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3301 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3361 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3421 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3481 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3541 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3601 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3661 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3721 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3781 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     3841 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3901 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3961 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4021 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4081 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4141 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4261 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4321 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4381 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4441 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4501 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4561 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4621 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4681 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4741 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4801 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4861 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4921 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     4981 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
