LOCUS       sequence-338254-        6955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mActZ-FY, a FiCE of iCMM, in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6955)
  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 6955)
  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-FY
FEATURES             Location/Qualifiers
     source          1..6955
                     /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"
     CDS             join(2905..2907,2908..3621)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2947..2968)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3208..3227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3555..3576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3743..3864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3780..3799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3834..3853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3871..4326)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4008..4029)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4220..4239
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4353..4457
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4459..4816
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4459..4650
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4482..4502)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4667..4802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4851..5645
                     /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             4851..5645
                     /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(4905..4924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5515..5534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5833..5852)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5877..5924
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6253..6841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6742..6761
                     /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 ggtcgtgagc aagggcgagg agctgttcac cggggtggtg
     2941 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3001 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3061 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     3121 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3181 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3241 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3301 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3361 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3421 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3481 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     3541 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3601 atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt
     3661 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3721 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3781 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3841 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3901 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3961 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4021 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4081 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4141 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4201 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4261 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4321 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4441 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4501 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4561 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4621 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4681 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4741 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4801 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4861 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4921 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4981 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     5041 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5101 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5161 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5221 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5281 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5341 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5401 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5461 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5521 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5581 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5641 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5701 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5761 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5821 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5881 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5941 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     6001 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6061 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6121 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6181 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6241 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6301 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6361 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6421 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6481 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6541 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6601 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6661 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6721 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6781 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6841 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6901 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
