LOCUS       sequence-260107-        5029 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 5029)
  TITLE     Okada Lab plasmids
REFERENCE   2  (bases 1 to 5029)
  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)-mNG
FEATURES             Location/Qualifiers
     source          1..5029
                     /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             925..1635
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1757..1878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1794..1813)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1848..1867
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1885..2340)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2022..2043)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2234..2253
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2367..2471
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2473..2830
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2473..2664
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2496..2516)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2681..2816
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2865..3659
                     /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             2865..3659
                     /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(2919..2938)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3529..3548
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3847..3866)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3891..3938
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4267..4855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4756..4775
                     /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 ggtcatggtg agcaagggcg aggaggataa catggcctct
      961 ctcccagcga cacatgagtt acacatcttt ggctccatca acggtgtgga ctttgacatg
     1021 gtgggtcagg gcaccggcaa tccaaatgat ggttatgagg agttaaacct gaagtccacc
     1081 aagggtgacc tccagttctc cccctggatt ctggtccctc atatcgggta tggcttccat
     1141 cagtacctgc cctaccctga cgggatgtcg cctttccagg ccgccatggt agatggctcc
     1201 ggataccaag tccatcgcac aatgcagttt gaagatggtg cctcccttac tgttaactac
     1261 cgctacacct acgagggaag ccacatcaaa ggagaggccc aggtgaaggg gactggtttc
     1321 cctgctgacg gtcctgtgat gaccaactcg ctgaccgctg cggactggtg caggtcgaag
     1381 aagacttacc ccaacgacaa aaccatcatc agtaccttta agtggagtta caccactgga
     1441 aatggcaagc gctaccggag cactgcgcgg accacctaca cctttgccaa gccaatggcg
     1501 gctaactatc tgaagaacca gccgatgtac gtgttccgta agacggagct caagcactcc
     1561 aagaccgagc tcaacttcaa ggagtggcaa aaggccttta ccgatgtgat gggcatggac
     1621 gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     1681 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     1741 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1801 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1861 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     1921 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1981 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2041 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2101 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2161 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2221 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2281 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2341 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2401 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2461 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     2521 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2581 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2641 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2701 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2761 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2821 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2881 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2941 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3001 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3061 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3121 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3181 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3241 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3301 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3361 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     3421 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3481 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3541 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3601 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3661 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3721 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3781 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     3841 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3901 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     3961 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4021 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4081 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4141 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4201 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4381 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4621 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4861 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4921 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     4981 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
