LOCUS       sequence-309482-        6981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MFN2:mcherry in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  AUTHORS   Bertolin G, Bulteau AL, Alves-Guerra MC, Burel A, Lavault MT, Gavard
            O, Le Bras S, Gagne JP, Poirier GG, Le Borgne R, Prigent C, Tramier 
            M
  TITLE     Aurora kinase A localises to mitochondria to control organelle 
            dynamics and energy production.
  JOURNAL   eLife. 2018 Aug 2;7. pii: 38111. doi: 10.7554/eLife.38111.
  PUBMED    30070631
REFERENCE   2  (bases 1 to 6981)
  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: "eLife."; 
            date: "2018-08-2"; volume: "7. pii: 38111. doi"; pages: " 
            10.7554/eLife.38111"
COMMENT     Sequence Label: pmCherry N1 MFN2
FEATURES             Location/Qualifiers
     source          1..6981
                     /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"
     RBS             1188..1196
                     /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)"
     regulatory      2566..2575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2572..3282
                     /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(2717..2735)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2985..3004
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3405..3526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3442..3461)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3496..3515
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3533..3988)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3670..3691)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3882..3901
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4015..4119
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4121..4478
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4121..4312
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4144..4164)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4329..4464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4513..5307
                     /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             4513..5307
                     /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(4567..4586)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5177..5196
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5495..5514)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5539..5586
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5915..6503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6404..6423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6678..6981
                     /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 actcagatct cgagctcaag cttcgaattc atgtccctgc tcttctctcg atgcaactct
      301 atcgtcacag tcaagaaaaa taagagacac atggctgagg tgaatgcatc cccacttaag
      361 cactttgtca ctgccaagaa gaagatcaat ggcatttttg agcagctggg ggcctacatc
      421 caggagagcg ccaccttcct tgaagacacg tacaggaatg cagaactgga ccccgttacc
      481 acagaagaac aggttctgga cgtcaaaggt tacctatcca aagtgagagg catcagtgag
      541 gtgctggctc ggaggcacat gaaagtggct ttttttggcc ggacgagcaa tgggaagagc
      601 accgtgatca atgccatgct ctgggacaaa gttctgccct ctgggattgg ccacaccacc
      661 aattgcttcc tgcgggtaga gggcacagat ggccatgagg cctttctcct taccgagggc
      721 tcagaggaaa agaggagtgc caagactgtg aaccagctgg cccatgccct ccaccaggac
      781 aagcagctcc atgccggcag cctagtgagt gtgatgtggc ccaactctaa gtgcccactt
      841 ctgaaggatg acctcgtttt gatggacagc cctggtattg atgtcaccac agagctggac
      901 agctggattg acaagttttg tctggatgct gatgtgtttg tgctggtggc caactcagag
      961 tccaccctga tgcagacgga aaagcacttc ttccacaagg tgagtgagcg tctctcccgg
     1021 ccaaacatct tcatcctgaa caaccgctgg gatgcatctg cctcagagcc cgagtacatg
     1081 gaggaggtgc ggcggcagca catggagcgt tgtaccagct tcctggtgga tgagctgggc
     1141 gtggtggatc gatcccaggc cggggaccgc atcttctttg tgtctgctaa ggaggtgctc
     1201 aacgccagga ttcagaaagc ccagggcatg cctgaaggag ggggcgctct cgcagaaggc
     1261 tttcaagtga ggatgtttga gtttcagaat tttgagagga gatttgagga gtgcatctcc
     1321 cagtctgcag tgaagaccaa gtttgagcag cacacggtcc gggccaagca gattgcagag
     1381 gcggttcgac tcatcatgga ctccctgcac atggcggctc gggagcagca ggtttactgc
     1441 gaggaaatgc gtgaagagcg gcaagaccga ctgaaattta ttgacaaaca gctggagctc
     1501 ttggctcaag actataagct gcgaattaag cagattacgg aggaagtgga gaggcaggtg
     1561 tcgactgcaa tggccgagga gatcaggcgc ctctctgtac tggtggacga ttaccagatg
     1621 gacttccacc cttctccagt agtcctcaag gtttataaga atgagctgca ccgccacata
     1681 gaggaaggac tgggtcgaaa catgtctgac cgctgctcca cggccatcac caactccctg
     1741 cagaccatgc agcaggacat gatagatggc ttgaaacccc tccttcctgt gtccgtgcgg
     1801 agtcagatag acatgctggt cccacgccag tgcttctccc tcaactatga cctaaactgt
     1861 gacaagctgt gtgctgactt ccaggaagac attgagttcc atttctctct cggatggacc
     1921 atgctggtga ataggttcct gggccccaag aacagccgtc gggccttgat gggctacaat
     1981 gaccaggtcc agcgtcccat ccctctgacg ccagccaacc ccagcatgcc cccactgcca
     2041 cagggctcgc tcacccagga ggagttcatg gtttccatgg ttaccggcct ggcctccttg
     2101 acatccagga cctccatggg cattcttgtt gttggaggag tggtgtggaa ggcagtgggc
     2161 tggcggctca ttgccctctc ctttgggctc tatggcctcc tctacgtcta tgagcgtctg
     2221 acctggacca ccaaggccaa ggagagggcc ttcaagcgcc agtttgtgga gcatgccagc
     2281 gagaagctgc agcttgtcat cagctacact ggctccaact gcagccacca agtccagcag
     2341 gaactgtctg ggacctttgc tcatctgtgt cagcaagttg acgtcacccg ggagaacctg
     2401 gagcaggaaa ttgccgccat gaacaagaaa attgaggttc ttgactcact tcagagcaaa
     2461 gcaaagctgc tcaggaataa agccggttgg ttggacagtg agctcaacat gttcacacac
     2521 cagtacctgc agcccagcag agcggatcca ggggatccac cggtcgccac catggtgagc
     2581 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     2641 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     2701 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     2761 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     2821 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     2881 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     2941 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     3001 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     3061 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     3121 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     3181 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     3241 ggccgccact ccaccggcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     3301 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3361 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3421 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3481 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3541 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3601 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3661 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3721 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3781 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3841 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3901 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3961 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4081 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4141 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4201 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4321 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4381 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4441 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4501 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4561 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4621 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4681 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4741 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4801 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4861 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4921 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4981 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5041 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5101 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5161 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5221 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5281 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5341 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5401 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5461 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5521 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5581 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5641 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5701 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5761 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5821 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5881 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5941 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6001 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6061 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6121 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6181 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6241 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6301 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6361 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6421 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6481 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6541 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6601 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
     6661 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6721 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6781 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6841 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6901 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6961 ttagtcatcg ctattaccat g
//
