LOCUS       sequence-353487-        7814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Drp1 Isoform 3 with K569A, K571A mutations in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7814)
  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 7814)
  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: 6xHis(TEV)-Drp1(K569A,K571A)-StrepII
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(327..350)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(2451..2471)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2472..2489)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2508..2530
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2545..2569
                     /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)."
     primer_bind     complement(2569..2588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2570..2588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2672..2691)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2897..2974
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2975..4057
                     /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     complement(2994..3013)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5369..5560
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5554..5576)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5662..5801
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5717..5736
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5816..5833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5987..6575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6067..6086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6746..7537,7538..7606))
                     /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     7369..7388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7607..7711)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7779..7797
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcttatttt tcgaactgag gatgagacca ccaaagatga
      361 gtctcccgga tttcagcaat aatttgactg gctccttgta atgcctttag catatcagct
      421 gcttctttcc tgcgctgtgc catgtcctca gattctgtca gaagatcatc caataaggat
      481 gatttataca gctggcctac tagctcactc tgaagagtgt ctttcacatg attaaccaaa
      541 aaatgcatta ctgcctttgg cacactgtct tgaatattct ttctgacaat gagaaaatat
      601 gatttaatga gtcgttcaat aacctcacaa tctcgctgtt cccgagcaga tagttttcgt
      661 gcaacaggaa ctggcacatc tagcaggttc acggcatgac ctttttgtgg actggctggc
      721 ataattggaa tgggtgctga tgcttcttct gctaataact cttcagcttt tgaagttttc
      781 agcattcctc tccagttgcc tgtggttggt tcttgaacac catctccaac cccaccacct
      841 ccagatgcaa ccttgtctcg tgatacagct gaaggtaatt ctctggctag cctgtttctc
      901 ctttgttcct ctatattatt gttcattagc ccacaagcat cagcaaagtc tggatgtttt
      961 gtgttgatat aagccagttc aattgccact aagttatgga ccatttcatt tgtaacaggc
     1021 aaccttttac gaagaagaca agtcaccact tcaactatgg catcatgaag tttaggaaat
     1081 cgtaacaatt cctgtgtact gtaattgcta cagtgctgaa tgatcctttg catttcctca
     1141 tgaaccagtt ccacacagcg gaggctgggc tcttctagac gtttgatttg ccgcttcacc
     1201 agtaactcaa atgaaacctc aggcacaaat aaagcaggac gaggaccagt agcatttcta
     1261 atggcagtca aaatgtcaat agtgttaagg ccaccaagtg gatcaacaga ttctaaggtt
     1321 cgcccaaaag tctcatggaa aatataacaa attctagcac caccgcatag ctccgaagtt
     1381 tcaatatatt ttgcagttcc ttcaatagtg ttacaatatt ctgtggcaaa tttggtaata
     1441 agttggagta aagtagcact tttatcatcc acgggttcac cgtagctatt tagaagagac
     1501 tgatactgag cagctagaac atttattctt gttttcaact ctggtaaaca atctctgatg
     1561 tgatgcatca gtaacctgtt tagagtccta gcaagatact ttgttccatt tctattggcc
     1621 agagatggat atttcttttg aagaaaagca tactcatcac ggattgaatc agttacactc
     1681 ttcttgttgt taatatctag ctggctcctg ttaactactc caattattcc aagtttgact
     1741 ggaataaccc ttcccatcaa tacatccatg gcatcagtac ccgcatccat gagatcaagt
     1801 ttagtgatta cagctagggt tctgcgacca tctggatcta cctctcttga aattttaagt
     1861 gcctctgatg ttgccatatc tgtattagca gcagtgacag cgaggataat ggaattagga
     1921 ttactgatga accgaagaat gagctctctg atttgaagct caatatcctt aggttgatca
     1981 cctacaggca ccttggtcat tcctggcaaa tccacaagtg tcaaattgac aacgttgggt
     2041 gaaaaaatct taagatgaat tggttcaggg cttactccct tattatttcc tgaaattctt
     2101 tctgtttcat tttcaatttc ttgtcgaatt tcatcaaaat ccgtgtaaag cttatttttg
     2161 gtgtgaagaa atttacccca ttcttctgct tccaccccat tttcttctcc tgttgttttc
     2221 cgtttatctt cttgtgaaac atggaccagt tgcagaatga gaggtctccg ggtgacaatt
     2281 ccagtacctc tgggaagcag gtccctcccc accaggcttt ctagcactga gctctttccg
     2341 ctgctctgcg ttcccactac gacgatttga ggcagctgga tgatgtcggc gcccaccgtg
     2401 ttgaagacgt cctggagctt gtttatgaca ggaattagcg cctccatcga gccttggaaa
     2461 tacagatttt cgtgatgatg atgatgatgg ctgctgccca tggtatatct ccttcttaaa
     2521 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
     2581 tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc
     2641 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     2701 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     2761 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     2821 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     2881 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     2941 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     3001 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     3061 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     3121 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     3181 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     3241 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     3301 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     3361 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     3421 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     3481 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     3541 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     3601 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     3661 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     3721 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     3781 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     3841 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     3901 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     3961 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4021 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     4081 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
     4141 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     4201 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
     4261 ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
     4321 tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
     4381 tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct
     4441 ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc
     4501 aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     4561 cggctcttac cagcctaact tcgatcattg gaccgctgat cgtcacggcg atttatgccg
     4621 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     4681 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     4741 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     4801 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     4861 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     4921 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     4981 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     5041 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     5101 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     5161 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     5221 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     5281 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     5341 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     5401 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     5461 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     5521 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5581 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5641 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     5701 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg
     5761 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     5821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6181 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6421 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6541 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6601 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6661 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6721 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6781 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6841 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6901 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6961 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7021 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     7081 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7141 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7201 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7261 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7321 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     7381 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7441 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7501 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7561 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7621 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7681 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7741 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7801 ttcgtcttca agaa
//
