LOCUS       sequence-309909-        6957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression and labeling mitofusin 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6957)
  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 6957)
  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: mCh-Mfn2
FEATURES             Location/Qualifiers
     source          1..6957
                     /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      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1260
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(698..716)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     966..985
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     RBS             2199..2207
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3722..3741)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3776..3795
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3813..4268)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3950..3971)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4162..4181
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4295..4399
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4401..4758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4401..4592
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4424..4444)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4609..4744
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4793..5587
                     /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             4793..5587
                     /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(4847..4866)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5457..5476
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5775..5794)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5819..5866
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6195..6783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6684..6703
                     /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 gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      601 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      661 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      721 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      781 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      901 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      961 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1021 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1081 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1141 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1201 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1261 tccggactca gatctcgagt gatgtccctg ctcttctctc gatgcaactc tatcgtcaca
     1321 gtcaagaaaa ataagagaca catggctgag gtgaatgcat ccccacttaa gcactttgtc
     1381 actgccaaga agaagatcaa tggcattttt gagcagctgg gggcctacat ccaggagagc
     1441 gccaccttcc ttgaagacac gtacaggaat gcagaactgg accccgttac cacagaagaa
     1501 caggttctgg acgtcaaagg ttacctatcc aaagtgagag gcatcagtga ggtgctggct
     1561 cggaggcaca tgaaagtggc tttttttggc cggacgagca atgggaagag caccgtgatc
     1621 aatgccatgc tctgggacaa agttctgccc tctgggattg gccacaccac caattgcttc
     1681 ctgcgggtag agggcacaga tggccatgag gcctttctcc ttaccgaggg ctcagaggaa
     1741 aagaggagtg ccaagactgt gaaccagctg gcccatgccc tccaccagga caagcagctc
     1801 catgccggca gcctagtgag tgtgatgtgg cccaactcta agtgcccact tctgaaggat
     1861 gacctcgttt tgatggacag ccctggtatt gatgtcacca cagagctgga cagctggatt
     1921 gacaagtttt gtctggatgc tgatgtgttt gtgctggtgg ccaactcaga gtccaccctg
     1981 atgcagacgg aaaagcactt cttccacaag gtgagtgagc gtctctcccg gccaaacatc
     2041 ttcatcctga acaaccgctg ggatgcatct gcctcagagc ccgagtacat ggaggaggtg
     2101 cggcggcagc acatggagcg ttgtaccagc ttcctggtgg atgagctggg cgtggtggat
     2161 cgatcccagg ccggggaccg catcttcttt gtgtctgcta aggaggtgct caacgccagg
     2221 attcagaaag cccagggcat gcctgaagga gggggcgctc tcgcagaagg ctttcaagtg
     2281 aggatgtttg agtttcagaa ttttgagagg agatttgagg agtgcatctc ccagtctgca
     2341 gtgaagacca agtttgagca gcacacggtc cgggccaagc agattgcaga ggcggttcga
     2401 ctcatcatgg actccctgca catggcggct cgggagcagc aggtttactg cgaggaaatg
     2461 cgtgaagagc ggcaagaccg actgaaattt attgacaaac agctggagct cttggctcaa
     2521 gactataagc tgcgaattaa gcagattacg gaggaagtgg agaggcaggt gtcgactgca
     2581 atggccgagg agatcaggcg cctctctgta ctggtggacg attaccagat ggacttccac
     2641 ccttctccag tagtcctcaa ggtttataag aatgagctgc accgccacat agaggaagga
     2701 ctgggtcgaa acatgtctga ccgctgctcc acggccatca ccaactccct gcagaccatg
     2761 cagcaggaca tgatagatgg cttgaaaccc ctccttcctg tgtctgtgcg gagtcagata
     2821 gacatgctgg tcccacgcca gtgcttctcc ctcaactatg acctaaactg tgacaagctg
     2881 tgtgctgact tccaggaaga cattgagttc catttctctc tcggatggac catgctggtg
     2941 aataggttcc tgggccccaa gaacagccgt cgggccttga tgggctacaa tgaccaggtc
     3001 cagcgtccca tccctctgac gccagccaac cccagcatgc ccccactgcc acagggctcg
     3061 ctcacccagg aggagttcat ggtttccatg gttaccggcc tggcctcctt gacatccagg
     3121 acctccatgg gcattcttgt tgttggagga gtggtgtgga aggcagtggg ctggcggctc
     3181 attgccctct cctttgggct ctatggcctc ctctacgtct atgagcgtct gacctggacc
     3241 accaaggcca aggagagggc cttcaagcgc cagtttgtgg agcatgccag cgagaagctg
     3301 cagcttgtca tcagctacac tggctccaac tgcagccacc aagtccagca ggaactgtct
     3361 gggacctttg ctcatctgtg tcagcaagtt gacgtcaccc gggagaacct ggagcaggaa
     3421 attgccgcca tgaacaagaa aattgaggtt cttgactcac ttcagagcaa agcaaagctg
     3481 ctcaggaata aagccggttg gttggacagt gagctcaaca tgttcacaca ccagtacctg
     3541 cagcccagca gatagggatc caccggatct agataactga tcataatcag ccataccaca
     3601 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3661 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3781 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     3841 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3901 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3961 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4021 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4081 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4141 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4201 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4261 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4321 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4381 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4441 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4501 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4561 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4621 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4681 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4741 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4801 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4861 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4921 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4981 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5041 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5101 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5161 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5221 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5281 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5341 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5401 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5461 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5521 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5581 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5761 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5821 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5881 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5941 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     6001 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     6901 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
