LOCUS       sequence-343849-        7814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Drp1 Isoform 3 with K557A, K560A 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(K557A,K560A)-StrepII
FEATURES             Location/Qualifiers
     source          1..7814
                     /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             2849..2866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2867..2887
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4988..5011
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5064..5082)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5078..5125
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5300..5305,5306..5322,5323..5328))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5355..5373)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5441..5545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5546..5614,5615..6406)
                     /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(5764..5783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6577..7165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7066..7085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7319..7336
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7351..7490
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7416..7435)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7576..7598
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7592..7783)
                     /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 tcctgcatta 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 gcagcagcca tcatcatcat catcacgaaa atctgtattt
     2881 ccaaggctcg atggaggcgc taattcctgt cataaacaag ctccaggacg tcttcaacac
     2941 ggtgggcgcc gacatcatcc agctgcctca aatcgtcgta gtgggaacgc agagcagcgg
     3001 aaagagctca gtgctagaaa gcctggtggg gagggacctg cttcccagag gtactggaat
     3061 tgtcacccgg agacctctca ttctgcaact ggtccatgtt tcacaagaag ataaacggaa
     3121 aacaacagga gaagaaaatg gggtggaagc agaagaatgg ggtaaatttc ttcacaccaa
     3181 aaataagctt tacacggatt ttgatgaaat tcgacaagaa attgaaaatg aaacagaaag
     3241 aatttcagga aataataagg gagtaagccc tgaaccaatt catcttaaga ttttttcacc
     3301 caacgttgtc aatttgacac ttgtggattt gccaggaatg accaaggtgc ctgtaggtga
     3361 tcaacctaag gatattgagc ttcaaatcag agagctcatt cttcggttca tcagtaatcc
     3421 taattccatt atcctcgctg tcactgctgc taatacagat atggcaacat cagaggcact
     3481 taaaatttca agagaggtag atccagatgg tcgcagaacc ctagctgtaa tcactaaact
     3541 tgatctcatg gatgcgggta ctgatgccat ggatgtattg atgggaaggg ttattccagt
     3601 caaacttgga ataattggag tagttaacag gagccagcta gatattaaca acaagaagag
     3661 tgtaactgat tcaatccgtg atgagtatgc ttttcttcaa aagaaatatc catctctggc
     3721 caatagaaat ggaacaaagt atcttgctag gactctaaac aggttactga tgcatcacat
     3781 cagagattgt ttaccagagt tgaaaacaag aataaatgtt ctagctgctc agtatcagtc
     3841 tcttctaaat agctacggtg aacccgtgga tgataaaagt gctactttac tccaacttat
     3901 taccaaattt gccacagaat attgtaacac tattgaagga actgcaaaat atattgaaac
     3961 ttcggagcta tgcggtggtg ctagaatttg ttatattttc catgagactt ttgggcgaac
     4021 cttagaatct gttgatccac ttggtggcct taacactatt gacattttga ctgccattag
     4081 aaatgctact ggtcctcgtc ctgctttatt tgtgcctgag gtttcatttg agttactggt
     4141 gaagcggcaa atcaaacgtc tagaagagcc cagcctccgc tgtgtggaac tggttcatga
     4201 ggaaatgcaa aggatcattc agcactgtag caattacagt acacaggaat tgttacgatt
     4261 tcctaaactt catgatgcca tagttgaagt ggtgacttgt cttcttcgta aaaggttgcc
     4321 tgttacaaat gaaatggtcc ataacttagt ggcaattgaa ctggcttata tcaacacaaa
     4381 acatccagac tttgctgatg cttgtgggct aatgaacaat aatatagagg aacaaaggag
     4441 aaacaggcta gccagagaat taccttcagc tgtatcacga gacaaggttg catctggagg
     4501 tggtggggtt ggagatggtg ttcaagaacc aaccacaggc aactggagag gaatgctggc
     4561 aacttcagca gctgaagagt tattagcaga agaaaaatca aaacccattc caattatgcc
     4621 agccagtcca caaaaaggtc atgccgtgaa cctgctagat gtgccagttc ctgttgcacg
     4681 aaaactatct gctcgggaac agcgagattg tgaggttatt gaacgactca ttaaatcata
     4741 ttttctcatt gtcagaaaga atattcaaga cagtgtgcca aaggcagtaa tgcatttttt
     4801 ggttaatcat gtgaaagaca ctcttcagag tgagctagta ggccagctgt ataaatcatc
     4861 cttattggat gatcttctga cagaatctga ggacatggca cagcgcagga aagaagcagc
     4921 tgatatgcta aaggcattac aaggagccag tcaaattatt gctgaaatcc gggagactca
     4981 tctttggtgg tctcatcctc agttcgaaaa ataagatccg gctgctaaca aagcccgaaa
     5041 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5101 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag
     5161 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg
     5221 aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt
     5281 aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaattc
     5341 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5401 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5461 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5521 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5581 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5641 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5701 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5761 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     5821 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5881 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5941 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6001 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6061 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     6121 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6181 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6241 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6301 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6361 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6421 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6481 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6541 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6601 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6661 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6721 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6781 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6841 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6901 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6961 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7021 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7081 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7141 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7201 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7261 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7321 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     7381 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7441 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     7501 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     7561 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     7621 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     7681 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     7741 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     7801 ggggatttct gttc
//
