LOCUS       sequence-343852-        7778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Drp1 Isoform 3 with A395D mutation in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7778)
  AUTHORS   Kamerkar SC, Kraus F, Sharpe AJ, Pucadyil TJ, Ryan MT
  TITLE     Dynamin-related protein 1 has membrane constricting and severing 
            abilities sufficient for mitochondrial and peroxisomal fission.
  JOURNAL   Nat Commun. 2018 Dec 7;9(1):5239. doi: 10.1038/s41467-018-07543-w.
  PUBMED    30531964
REFERENCE   2  (bases 1 to 7778)
  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: "Nat 
            Commun."; date: "2018-12-7"; volume: "9(1):5239. doi"; pages: " 
            10.1038/s41467-018-07543-w"
COMMENT     Sequence Label: Drp1(A395D)-StrepII
FEATURES             Location/Qualifiers
     source          1..7778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1281..2363)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2325..2344
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2364..2441)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2647..2666
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2750..2769
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2750..2768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2769..2793
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2808..2830
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4952..4975
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5028..5046)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5042..5089
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5264..5269,5270..5286,5287..5292))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5319..5337)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5405..5509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5510..5578,5579..6370)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5728..5747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6541..7129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7030..7049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7283..7300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7315..7454
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7380..7399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7540..7562
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7556..7747)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
       61 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
      121 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
      181 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
      241 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
      301 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
      361 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
      421 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
      481 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
      541 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
      601 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
      661 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
      721 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
      781 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
      841 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
      901 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
      961 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     1021 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     1081 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     1141 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     1201 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     1261 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1321 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     1381 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     1441 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     1501 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     1561 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     1621 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     1681 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     1741 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     1801 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     1861 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     1921 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     1981 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     2041 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     2101 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     2161 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     2221 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     2281 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     2341 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     2401 catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc
     2461 cttatgcgac tcccgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     2521 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     2581 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     2641 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     2701 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     2761 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
     2821 aagaaggaga tataccatgg gcagcagcgg ctcgatggag gcgctaattc ctgtcataaa
     2881 caagctccag gacgtcttca acacggtggg cgccgacatc atccagctgc ctcaaatcgt
     2941 cgtagtggga acgcagagca gcggaaagag ctcagtgcta gaaagcctgg tggggaggga
     3001 cctgcttccc agaggtactg gaattgtcac ccggagacct ctcattctgc aactggtcca
     3061 tgtttcacaa gaagataaac ggaaaacaac aggagaagaa aatggggtgg aagcagaaga
     3121 atggggtaaa tttcttcaca ccaaaaataa gctttacacg gattttgatg aaattcgaca
     3181 agaaattgaa aatgaaacag aaagaatttc aggaaataat aagggagtaa gccctgaacc
     3241 aattcatctt aagatttttt cacccaacgt tgtcaatttg acacttgtgg atttgccagg
     3301 aatgaccaag gtgcctgtag gtgatcaacc taaggatatt gagcttcaaa tcagagagct
     3361 cattcttcgg ttcatcagta atcctaattc cattatcctc gctgtcactg ctgctaatac
     3421 agatatggca acatcagagg cacttaaaat ttcaagagag gtagatccag atggtcgcag
     3481 aaccctagct gtaatcacta aacttgatct catggatgcg ggtactgatg ccatggatgt
     3541 attgatggga agggttattc cagtcaaact tggaataatt ggagtagtta acaggagcca
     3601 gctagatatt aacaacaaga agagtgtaac tgattcaatc cgtgatgagt atgcttttct
     3661 tcaaaagaaa tatccatctc tggccaatag aaatggaaca aagtatcttg ctaggactct
     3721 aaacaggtta ctgatgcatc acatcagaga ttgtttacca gagttgaaaa caagaataaa
     3781 tgttctagct gctcagtatc agtctcttct aaatagctac ggtgaacccg tggatgataa
     3841 aagtgctact ttactccaac ttattaccaa atttgccaca gaatattgta acactattga
     3901 aggaactgca aaatatattg aaacttcgga gctatgcggt ggtgctagaa tttgttatat
     3961 tttccatgag acttttgggc gaaccttaga atctgttgat ccacttggtg gccttaacac
     4021 tattgacatt ttgactgaca ttagaaatgc tactggtcct cgtcctgctt tatttgtgcc
     4081 tgaggtttca tttgagttac tggtgaagcg gcaaatcaaa cgtctagaag agcccagcct
     4141 ccgctgtgtg gaactggttc atgaggaaat gcaaaggatc attcagcact gtagcaatta
     4201 cagtacacag gaattgttac gatttcctaa acttcatgat gccatagttg aagtggtgac
     4261 ttgtcttctt cgtaaaaggt tgcctgttac aaatgaaatg gtccataact tagtggcaat
     4321 tgaactggct tatatcaaca caaaacatcc agactttgct gatgcttgtg ggctaatgaa
     4381 caataatata gaggaacaaa ggagaaacag gctagccaga gaattacctt cagctgtatc
     4441 acgagacaag gttgcatctg gaggtggtgg ggttggagat ggtgttcaag aaccaaccac
     4501 aggcaactgg agaggaatgc tgaaaacttc aaaagctgaa gagttattag cagaagaaaa
     4561 atcaaaaccc attccaatta tgccagccag tccacaaaaa ggtcatgccg tgaacctgct
     4621 agatgtgcca gttcctgttg cacgaaaact atctgctcgg gaacagcgag attgtgaggt
     4681 tattgaacga ctcattaaat catattttct cattgtcaga aagaatattc aagacagtgt
     4741 gccaaaggca gtaatgcatt ttttggttaa tcatgtgaaa gacactcttc agagtgagct
     4801 agtaggccag ctgtataaat catccttatt ggatgatctt ctgacagaat ctgaggacat
     4861 ggcacagcgc aggaaagaag cagctgatat gctaaaggca ttacaaggag ccagtcaaat
     4921 tattgctgaa atccgggaga ctcatctttg gtggtctcat cctcagttcg aaaaataaga
     4981 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     5041 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     5101 aactatatcc ggatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct
     5161 acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac
     5221 ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
     5281 ataagctgtc aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt
     5341 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5521 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5581 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5641 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5701 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     5761 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5821 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5881 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5941 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6001 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     6061 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6121 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6181 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6241 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6301 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6361 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6421 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6481 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6661 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7201 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7261 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     7321 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     7381 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     7441 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     7501 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     7561 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     7621 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     7681 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     7741 tggtcactga tgcctccgtg taagggggat ttctgttc
//
