LOCUS       sequence-338252-        6956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mActZ-FmCh, a FiCE of iCMM, in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  AUTHORS   Miyamoto T, Uosaki H, Mizunoe Y, Han SI, Goto S, Yamanaka D, Masuda 
            M, Yoneyama Y, Nakamura H, Hattori N, Takeuchi Y, Ohno H, Sekiya M, 
            Matsuzaka T, Hakuno F, Takahashi SI, Yahagi N, Ito K, Shimano H
  TITLE     Rapid manipulation of mitochondrial morphology in a living cell with
            iCMM.
  JOURNAL   Cell Rep Methods. 2021 Jul 22;1(4):100052. doi: 
            10.1016/j.crmeth.2021.100052. eCollection 2021 Aug 23.
  PUBMED    35475143
REFERENCE   2  (bases 1 to 6956)
  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: "Cell Rep 
            Methods."; date: "2021-07-22"; volume: "1(4):100052. doi"; pages: " 
            10.1016/j.crmeth.2021.100052. eCollection 2021 Aug 23"
COMMENT     Sequence Label: mActZ-FmCh
FEATURES             Location/Qualifiers
     source          1..6956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2563..2883
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     regulatory      2905..2914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2911..3621
                     /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(3056..3074)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3324..3343
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3744..3865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3781..3800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3835..3854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3872..4327)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4009..4030)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4221..4240
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4354..4458
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4460..4817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4460..4651
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4483..4503)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4668..4803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4852..5646
                     /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             4852..5646
                     /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(4906..4925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5516..5535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5834..5853)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5878..5925
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6254..6842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6743..6762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatggct acagattcag aggatagctc cctgaacaca
      661 gacgagtggg aggaggagaa gaccgaggag cagccctccg aagtgaatac aggccctaga
      721 tacgagaccg ccagagaggt gtctagccgg gacatcaagg agctggagaa gtctaacaag
      781 gtgcgcaaca caaataaggc cgatctgatc gccatgctga aggagaaggc cgagaagggc
      841 ccaaatatca acaataacaa ttccgagcag accgagaacg ccgccatcaa tgaggaggcc
      901 tctggagcag acaggccagc aatccaggtg gagaggagac acccaggact gccatccgat
      961 tctgccgcag agatcaagaa gcggcgcaag gccatcgcct cctctgactc tgagctggag
     1021 agcctgacat atcctgataa gccaaccaag gtgaacaaga agaaggtggc aaaggagagc
     1081 gtggcagacg ccagcgagtc cgacctggat agctccatgc agtccgccga tgagtctagc
     1141 ccacagcccc tgaaggccaa ccagcagccc ttctttccta aggtgttcaa gaagatcaag
     1201 gacgccggca agtgggtgcg cgacaagatc gatgagaatc ccgaggtgaa gaaggccatc
     1261 gtggacaaga gcgccggcct gatcgatcag ctgctgacca agaagaagag cgaggaggtg
     1321 aacgccgcgg ccccccgccc gtctcccgcg atctccgttt cggtctcggc tccggctttt
     1381 tacgccccgc agaagaagtt cggccctgtg gtggccccaa agcccaaagt gaatcccttc
     1441 cggcccgggg acagcgagcc tcccccggca cccggggccc agcgcgcaca gatgggccgg
     1501 gtgggcgaga ttcccccgcc gcccccggaa gactttcccc tgcctccacc tccccttgct
     1561 ggggatggcg acgatgcaga gggtgctctg ggaggtgcct tcccgccgcc ccctcccccg
     1621 atcgaggaat catttccccc tgcgcctctg gaggaggaga tcttcccttc cccgccgcct
     1681 cctccggagg aggagggagg gcctgaggcc cccataccgc ccccaccaca gcccagggag
     1741 aaggtgagca gtattgattt ggagatcgac tctctgtcct cactgctgga tgacatgacc
     1801 aagaatgatc ctttcaaagc ccgggtgtca tctggatatg tgcccccacc agtggccact
     1861 ccattcagtt ccaagtccag taccaagcct gcagccgggg gcacagcacc cctgcctcct
     1921 tggaagtccc cttccagctc ccagcctctg ccccaggttc cggctccggc tcagagccag
     1981 acacagttcc atgttcagcc ccagccccag cccaagcctc aggtccaact ccatgtccag
     2041 tcccagaccc agcctgtgtc tttggctaac acccagcccc gagggccccc agcctcatct
     2101 ccggctccag cccctaagtt ttctccagtg actcctaagt ttactcctgt ggcttccaag
     2161 ttcagtcctg gagccccagg tggatctggg tcacaaccaa atcaaaaatt ggggcacccc
     2221 gaagctcttt ctgctggcac aggctcccct caacctccca gcttcaccta tgcccagcag
     2281 agggagaagc cccgagtgca ggagaagcag caccccgtgc ccccaccggc tcagaaccaa
     2341 aaccaggtgc gctcccctgg ggccccaggg cccctgactc tgaaggaggt ggaggagctg
     2401 gagcagctga cccagcagct aatgcaggac atggagcatc ctcagaggca gaatgtggct
     2461 gtccgaattc tgcagtcgac ggtaccgcgg gcccgggatc caagtgctgg tggtagtgct
     2521 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtggagtgca ggtggaaacc
     2581 atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac
     2641 accgggatgc ttgaagatgg aaagaaattt gattcctccc gggacagaaa caagcccttt
     2701 aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg
     2761 agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg tgccactggg
     2821 cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg
     2881 gaaagtgctg gtggtccacc ggtcgccacc atggtaagca agggcgagga ggataacatg
     2941 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     3001 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3061 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     3121 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     3181 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     3241 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     3301 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3361 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     3421 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     3481 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     3541 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     3601 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     3661 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3721 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3781 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3841 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3901 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3961 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4021 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4081 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4141 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4201 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4261 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4321 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4441 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4501 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4561 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4621 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4681 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4741 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4801 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4861 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4921 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4981 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     5041 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5101 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5161 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5221 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5281 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5341 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5401 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5461 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5521 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5581 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5641 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5701 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5761 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5821 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5881 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5941 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6001 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6061 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6121 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6181 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
