LOCUS       sequence-462165-        6783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses K38A variant of human DRP1 with mCherry tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6783)
  AUTHORS   Begeman A, Smolka JA, Shami A, Waingankar TP, Lewis SC
  TITLE     A spatial atlas of mitochondrial gene expression reveals dynamic 
            translation hubs and remodeling in stress.
  JOURNAL   bioRxiv [Preprint]. 2024 Aug 17:2024.08.05.604215. doi: 
            10.1101/2024.08.05.604215.
  PUBMED    39149346
REFERENCE   2  (bases 1 to 6783)
  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: "bioRxiv 
            [Preprint]. 2024 Aug 17:2024.08.05.604215. doi: 
            10.1101/2024.08.05.604215."
COMMENT     Sequence Label: mCherry-DRP1 K38A
FEATURES             Location/Qualifiers
     source          1..6783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1316..2023
                     /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(1461..1479)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1729..1748
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4274..4395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4311..4330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4365..4384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4402..4857)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4539..4560)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4751..4770
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4884..4988
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4990..5347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4990..5181
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5013..5033)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5198..5333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5382..6176
                     /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             5382..6176
                     /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(5436..5455)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6046..6065
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6364..6383)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6408..6455
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtatcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     1381 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1441 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1501 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     1561 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1621 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1681 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1741 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1801 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1861 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1921 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1981 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctcg
     2041 agtgatggag gcgctaattc ctgtcataaa caagctccag gacgtcttca acacggtggg
     2101 cgccgacatc atccagctgc ctcaaatcgt cgtagtggga acgcagagca gcggagccag
     2161 ctcagtgcta gaaagcctgg tggggaggga cctgcttccc agaggtactg gaattgtcac
     2221 ccggagacct ctcattctgc aactggtcca tgtttcacaa gaagataaac ggaaaacaac
     2281 aggagaagaa aatggagtgg aagcagaaga atggggtaaa tttcttcaca ccaaaaataa
     2341 gctttacacg gattttgatg aaattcgaca agaaattgaa aatgaaacag aaagaatttc
     2401 aggaaataat aagggagtaa gccctgaacc aattcatctt aagatttttt cacccaacgt
     2461 tgtcaatttg acacttgtgg atttgccagg aatgaccaag gtgcctgtag gtgatcaacc
     2521 taaggatatt gagcttcaaa tcagagagct cattcttcgg ttcatcagta atcctaattc
     2581 cattatcctc gctgtcactg ctgctaatac agatatggca acatcagagg cacttaaaat
     2641 ttcaagagag gtagatccag atggtcgcag aaccctagct gtaatcacta aacttgatct
     2701 catggatgcg ggtactgatg ccatggatgt attgatggga agggttattc cagtcaaact
     2761 tggaataatt ggagtagtta acaggagcca gctagatatt aacaacaaga agagtgtaac
     2821 tgattcaatc cgtgatgagt atgcttttct tcaaaagaaa tatccatctc tggccaatag
     2881 aaatggaaca aagtatcttg ctaggactct aaacaggtta ctgatgcatc acatcagaga
     2941 ttgtttacca gagttgaaaa cgagaataaa tgttctagct gctcagtatc agtctcttct
     3001 aaatagctac ggtgaacccg tggatgataa aagtgctact ttactccaac ttattaccaa
     3061 atttgccaca gaatattgta acactattga aggaactgca aaatatattg aaacttcgga
     3121 gctatgcggt ggtgctagaa tttgttatat tttccatgag acttttgggc gaaccttaga
     3181 atctgttgat ccacttggtg gccttaacac tattgacatt ttgactgcca ttagaaatgc
     3241 tactggtcct cgtcctgctt tatttgtgcc tgaggtttca tttgagttac tggtgaagcg
     3301 gcaaatcaaa cgtctagaag agcccagcct ccgctgtgtg gaactggttc atgaggaaat
     3361 gcaaaggatc attcagcact gtagcaatta cagtacacag gaattgttac gatttcctaa
     3421 acttcatgat gccatagttg aagtggtgac ttgtcttctt cgtaaaaggt tgcctgttac
     3481 aaatgaaatg gtccataact tagtggcaat tgaactggct tatatcaaca caaaacatcc
     3541 agactttgct gatgcttgtg ggctaatgaa caataatata gaggaacaaa ggagaaacag
     3601 gctagccaga gaattacctt cagctgtatc acgagacaag gttgcatctg gaggtggtgg
     3661 ggttggagat ggtgttcaag aaccaaccac aggcaactgg agaggaatgc tgaaaacttc
     3721 aaaagctgaa gagttattag cagaagaaaa atcaaaaccc attccaatta tgccagccag
     3781 tccacaaaaa ggtcatgccg tgaacctgct agatgtgcca gttcctgttg cacgaaaact
     3841 atctgctcgg gaacagcgag attgtgaggt tattgaacga ctcattaaat catattttct
     3901 cattgtcaga aagaatattc aagacagtgt gccaaaggca gtaatgcatt ttttggttaa
     3961 tcatgtgaaa gacactcttc agagtgagct agtaggccag ctgtataaat catccttatt
     4021 ggatgatctt ctgacagaat ctgaggacat ggcacagcgc aggaaagaag cagctgatat
     4081 gctaaaggca ttacaaggag ccagtcaaat tattgctgaa atccgggaga ctcatctttg
     4141 gtgaggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     4201 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4261 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4321 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4381 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4441 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4501 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4561 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4621 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4681 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4741 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4801 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4861 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4921 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4981 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     5041 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5101 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5161 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5221 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5281 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5341 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5401 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5461 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5521 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5581 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5641 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5701 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5761 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5821 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5881 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5941 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6001 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6061 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6121 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6181 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6241 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6301 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6361 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6421 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6481 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6541 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6601 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6661 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6721 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6781 ttc
//
