LOCUS       sequence-309902-        4647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent labeling of the outer mitochondrial membrane (OMM) in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4647)
  AUTHORS   Abrisch RG, Gumbin SC, Wisniewski BT, Lackner LL, Voeltz GK
  TITLE     Fission and fusion machineries converge at ER contact sites to 
            regulate mitochondrial morphology.
  JOURNAL   J Cell Biol. 2020 Apr 6;219(4). pii: 133843. doi: 
            10.1083/jcb.201911122.
  PUBMED    32328629
REFERENCE   2  (bases 1 to 4647)
  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: "J Cell 
            Biol."; date: "2020-04-6"; volume: "219(4). pii: 133843. doi"; 
            pages: " 10.1083/jcb.201911122"
COMMENT     Sequence Label: SNAP-OMP25
FEATURES             Location/Qualifiers
     source          1..4647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             249..794
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    1071..1192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1108..1127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1162..1181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1199..1654)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1336..1357)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1548..1567
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1681..1785
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1787..2144
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1787..1978
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1810..1830)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1995..2130
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2179..2973
                     /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             2179..2973
                     /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(2233..2252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2843..2862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3161..3180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3205..3252
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3581..4169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4070..4089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4344..4647
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
      301 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
      361 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
      421 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
      481 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
      541 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
      601 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
      661 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac gtggggggct
      721 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
      781 agcctgggct gggtagcggt agcggtggca gctctcgagg tacgcgtggg cgaggcgacg
      841 gagagccgag tggagttcct gtagctgtgg tgctgctgcc agtgtttgcc cttaccctgg
      901 tagcagtttg ggccttcgtg agataccgaa agcagctctg aggatccacc ggatctagat
      961 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1021 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1081 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1141 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     1201 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1261 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1321 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1381 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     1441 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     1501 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     1561 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     1621 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     1681 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1741 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     1801 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1861 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1921 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1981 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2041 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2101 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2161 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2221 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2281 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2341 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2401 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2461 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2521 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2581 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2641 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2701 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2761 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2821 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2881 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2941 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3001 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3061 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3121 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3181 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3241 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3301 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3361 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     3421 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3481 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3541 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3601 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3661 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3721 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3781 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3841 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3901 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3961 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4021 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4081 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4141 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4201 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4261 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     4321 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     4381 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     4441 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     4501 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     4561 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     4621 tacgtattag tcatcgctat taccatg
//
