LOCUS       sequence-309911-        6627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression and labeling of Drp1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6627)
  AUTHORS   Abrisch RG, Gumbin SC, Wisniewski BT, Lackner LL, Voeltz GK
  TITLE     Fission and fusion machineries converge at ER contact sites to 
            regulate mitochondrial morphology.
  JOURNAL   J Cell Biol. 2020 Apr 6;219(4). pii: 133843. doi: 
            10.1083/jcb.201911122.
  PUBMED    32328629
REFERENCE   2  (bases 1 to 6627)
  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: "J Cell 
            Biol."; date: "2020-04-6"; volume: "219(4). pii: 133843. doi"; 
            pages: " 10.1083/jcb.201911122"
COMMENT     Sequence Label: SNAP-Drp1
FEATURES             Location/Qualifiers
     source          1..6627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1098
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    3355..3476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3392..3411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3446..3465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3483..3938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3620..3641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3832..3851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3965..4069
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4071..4428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4071..4262
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4094..4114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4279..4414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4463..5257
                     /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             4463..5257
                     /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(4517..4536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5127..5146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5445..5464)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5489..5536
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5865..6453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6354..6373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggacaa agactgcgaa atgaagcgca ccaccctgga tagccctctg
      601 ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc gtatcatctt cctgggcaaa
      661 ggaacatctg ccgccgacgc cgtggaagtg cctgccccag ccgccgtgct gggcggacca
      721 gagccactga tgcaggccac cgcctggctc aacgcctact ttcaccagcc tgaggccatc
      781 gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc agcaggagag ctttacccgc
      841 caggtgctgt ggaaactgct gaaagtggtg aagttcggag aggtcatcag ctacagccac
      901 ctggccgccc tggccggcaa tcccgccgcc accgccgccg tgaaaaccgc cctgagcgga
      961 aatcccgtgc ccattctgat cccctgccac cgggtggtgc agggcgacct ggacgtgggg
     1021 ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg cccacgaggg ccacagactg
     1081 ggcaagcctg ggctgggtag cggtagcggt ggcagctctc gagtgatgga ggcgctaatt
     1141 cctgtcataa acaagctcca ggacgtcttc aacacggtgg gcgccgacat catccagctg
     1201 cctcaaatcg tcgtagtggg aacgcagagc agcggaaaga gctccgtgct agaaagcctg
     1261 gtggggaggg acctgcttcc cagaggtact ggaattgtca cccggagacc tctcattctg
     1321 caactggtcc atgtttcaca agaagataaa cggaaaacaa caggagaaga aaatggagtg
     1381 gaagcagaag aatggggtaa atttcttcac accaaaaata agctttacac ggattttgat
     1441 gaaattcgac aagaaattga aaatgaaaca gaaagaattt caggaaataa taagggagta
     1501 agccctgaac caattcatct taagattttt tcacccaacg ttgtcaattt gacacttgtg
     1561 gatttgccag gaatgaccaa ggtgcctgta ggtgatcaac ctaaggatat tgagcttcaa
     1621 atcagagagc tcattcttcg gttcatcagt aatcctaatt ccattatcct cgctgtcact
     1681 gctgctaata cagatatggc aacatcagag gcacttaaaa tttcaagaga ggtagatcca
     1741 gatggtcgca gaaccctagc tgtaatcact aaacttgatc tcatggatgc gggtactgat
     1801 gccatggatg tattgatggg aagggttatt ccagtcaaac ttggaataat tggagtagtt
     1861 aacaggagcc agctagatat taacaacaag aagagtgtaa ctgattcaat ccgtgatgag
     1921 tatgcttttc ttcaaaagaa atatccatct ctggccaata gaaatggaac aaagtatctt
     1981 gctaggactc taaacaggtt actgatgcat cacatcagag attgtttacc agagttgaaa
     2041 acgagaataa atgttctagc tgctcagtat cagtctcttc taaatagcta cggtgaaccc
     2101 gtggatgata aaagtgctac tttactccaa cttattacca aatttgccac agaatattgt
     2161 aacactattg aaggaactgc aaaatatatt gaaacttcgg agctatgcgg tggtgctaga
     2221 atttgttata ttttccatga gacttttggg cgaaccttag aatctgttga tccacttggt
     2281 ggccttaaca ctattgacat tttgactgcc attagaaatg ctactggtcc tcgtcctgct
     2341 ttatttgtgc ctgaggtttc atttgagtta ctggtgaagc ggcaaatcaa acgtctagaa
     2401 gagcccagcc tccgctgtgt ggaactggtt catgaggaaa tgcaaaggat cattcagcac
     2461 tgtagcaatt acagtacaca ggaattgtta cgatttccta aacttcatga tgccatagtt
     2521 gaagtggtga cttgtcttct tcgtaaaagg ttgcctgtta caaatgaaat ggtccataac
     2581 ttagtggcaa ttgaactggc ttatatcaac acaaaacatc cagactttgc tgatgcttgt
     2641 gggctaatga acaataatat agaggaacaa aggagaaaca ggctagccag agaattacct
     2701 tcagctgtat cacgagacaa ggttgcatct ggaggtggtg gggttggaga tggtgttcaa
     2761 gaaccaacca caggcaactg gagaggaatg ctgaaaactt caaaagctga agagttatta
     2821 gcagaagaaa aatcaaaacc cattccaatt atgccagcca gtccacaaaa aggtcatgcc
     2881 gtgaacctgc tagatgtgcc agttcctgtt gcacgaaaac tatctgctcg ggaacagcga
     2941 gattgtgagg ttattgaacg actcattaaa tcatattttc tcattgtcag aaagaatatt
     3001 caagacagtg tgccaaaggc agtaatgcat tttttggtta atcatgtgaa agacactctt
     3061 cagagtgagc tagtaggcca gctgtataaa tcatccttat tggatgatct tctgacagaa
     3121 tctgaggaca tggcacagcg caggaaagaa gcagctgata tgctaaaggc attacaagga
     3181 gccagtcaaa ttattgctga aatccgggag actcatcttt ggtgaggatc caccggatct
     3241 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3301 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3481 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3541 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3601 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3661 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3721 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3781 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3841 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3901 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4081 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4261 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4321 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4381 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4441 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4501 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4561 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4621 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4681 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4741 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4801 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4861 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4921 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4981 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5041 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5101 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5161 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5221 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5281 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5341 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5401 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5461 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5521 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5581 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5641 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5701 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6541 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6601 agttcatagc ccatatatgg agttccg
//
