LOCUS       sequence-304569-        5727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mcherry:MFF in cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5727)
  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 5727)
  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 C1 MFF
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     276..295
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(594..613)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    638..685
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1014..1602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1503..1522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1777..2080
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2081..2284
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2235..2255
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3392..3401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3398..4108
                     /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(3543..3561)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3811..3830
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4231..4352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4268..4287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4322..4341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4359..4814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4496..4517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4708..4727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4841..4945
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4947..5304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4947..5138
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4970..4990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5155..5290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5339..5727,1..406)
                     /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             join(5339..5727,1..406)
                     /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(5393..5412)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
       61 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
      121 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
      181 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
      241 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
      301 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
      361 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
      421 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
      481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
      541 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
      601 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
      661 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
      721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
      781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
      841 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
      901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1141 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1681 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     1741 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     1801 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     1861 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1921 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     1981 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2041 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2101 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2161 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2221 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2281 agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag
     2341 cttcgaattc atgagtaaag gaacaagcag tgacacatca ctaggaaggg tgagcagggc
     2401 agcatttcct tctcccactg ctgctgagat ggcagaaatt agtcgaattc agtacgaaat
     2461 ggaatatact gaaggcatta gtcagcgaat gagggtccca gaaaagttaa aagtagcacc
     2521 gccaaacgct gacctggaac aaggattcca agaaggagtt ccaaatgcta gtgtgataat
     2581 gcaagttccg gagaggattg ttgtagcagg aaataatgaa gatgtttcat tttcaagacc
     2641 agcagatctt gaccttattc agtcaactcc ctttaaaccc ctggcactga aaacaccacc
     2701 tcgtgtactt acgctgagtg aaagaccact agattttctg gatttagaaa gacctcctac
     2761 aacccctcaa aatgaagaaa tccgagcagt tggcagacta aaaagagagc ggtctatgag
     2821 tgaaaatgct gttcgccaaa atggacagct ggtcagaaat gattctctgt ggcacagatc
     2881 agattctgcc ccaagaaata aaatttcaag gttccaggca ccgatttctg caccggagta
     2941 cactgtgaca ccatcgccac aacaggctcg ggtctgtcct ccccatatgt tacctgaaga
     3001 tggagctaat ctttcctctg ctcgtggcat tttgtcgctt atccagtctt ctactcgtag
     3061 ggcataccag cagatcttgg atgtgctgga tgaaaatcgc agacctgtgt tgcgtggtgg
     3121 gtctgctgcc gccacttcta atcctcatca tgacaacgtc aggtatggca tttcaaatat
     3181 agatacaacc attgaaggaa cgtcagatga cctgactgtt gtagatgcag cttcactaag
     3241 acgacagata atcaaactaa atagacgtct acaacttctg gaagaggaga acaaagaacg
     3301 tgctaaaaga gaaatggtca tgtattcaat tactgtagct ttctggctgc ttaatagctg
     3361 gctctggttt cgccgcgggg atccaccggt cgccaccatg gtgagcaagg gcgaggagga
     3421 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     3481 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     3541 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     3601 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     3661 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     3721 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     3781 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     3841 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     3901 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     3961 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     4021 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     4081 cggcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     4141 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4201 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4261 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4321 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     4381 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4441 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4501 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4561 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4621 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4681 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4741 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4801 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4861 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4921 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4981 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5101 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5161 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5221 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5281 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5341 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5401 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5461 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5521 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5581 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5641 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5701 gcggctgcat acgcttgatc cggctac
//
