LOCUS       sequence-343837-        7814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Drp1 Isoform 3 with K38A mutation 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(K38A)-StrepII
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     897..916
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(936..1013)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1219..1238
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1322..1341
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1322..1340
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1341..1365
                     /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             1380..1402
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1421..1438
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1439..1459
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3560..3583
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3636..3654)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3650..3697
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3872..3877,3878..3894,3895..3900))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3927..3945)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4013..4117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4118..4186,4187..4978)
                     /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(4336..4355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5149..5737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5638..5657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5891..5908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5923..6062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5988..6007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6148..6170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6164..6355)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7667..7814,1..935))
                     /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"
ORIGIN
        1 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
       61 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
      121 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
      181 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
      241 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
      301 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
      361 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
      421 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
      481 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
      541 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
      601 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
      661 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
      721 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
      781 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
      841 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
      901 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
      961 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     1021 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     1081 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     1141 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     1201 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     1261 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     1321 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
     1381 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacga
     1441 aaatctgtat ttccaaggct cgatggaggc gctaattcct gtcataaaca agctccagga
     1501 cgtcttcaac acggtgggcg ccgacatcat ccagctgcct caaatcgtcg tagtgggaac
     1561 gcagagcagc ggagcgagct cagtgctaga aagcctggtg gggagggacc tgcttcccag
     1621 aggtactgga attgtcaccc ggagacctct cattctgcaa ctggtccatg tttcacaaga
     1681 agataaacgg aaaacaacag gagaagaaaa tggggtggaa gcagaagaat ggggtaaatt
     1741 tcttcacacc aaaaataagc tttacacgga ttttgatgaa attcgacaag aaattgaaaa
     1801 tgaaacagaa agaatttcag gaaataataa gggagtaagc cctgaaccaa ttcatcttaa
     1861 gattttttca cccaacgttg tcaatttgac acttgtggat ttgccaggaa tgaccaaggt
     1921 gcctgtaggt gatcaaccta aggatattga gcttcaaatc agagagctca ttcttcggtt
     1981 catcagtaat cctaattcca ttatcctcgc tgtcactgct gctaatacag atatggcaac
     2041 atcagaggca cttaaaattt caagagaggt agatccagat ggtcgcagaa ccctagctgt
     2101 aatcactaaa cttgatctca tggatgcggg tactgatgcc atggatgtat tgatgggaag
     2161 ggttattcca gtcaaacttg gaataattgg agtagttaac aggagccagc tagatattaa
     2221 caacaagaag agtgtaactg attcaatccg tgatgagtat gcttttcttc aaaagaaata
     2281 tccatctctg gccaatagaa atggaacaaa gtatcttgct aggactctaa acaggttact
     2341 gatgcatcac atcagagatt gtttaccaga gttgaaaaca agaataaatg ttctagctgc
     2401 tcagtatcag tctcttctaa atagctacgg tgaacccgtg gatgataaaa gtgctacttt
     2461 actccaactt attaccaaat ttgccacaga atattgtaac actattgaag gaactgcaaa
     2521 atatattgaa acttcggagc tatgcggtgg tgctagaatt tgttatattt tccatgagac
     2581 ttttgggcga accttagaat ctgttgatcc acttggtggc cttaacacta ttgacatttt
     2641 gactgccatt agaaatgcta ctggtcctcg tcctgcttta tttgtgcctg aggtttcatt
     2701 tgagttactg gtgaagcggc aaatcaaacg tctagaagag cccagcctcc gctgtgtgga
     2761 actggttcat gaggaaatgc aaaggatcat tcagcactgt agcaattaca gtacacagga
     2821 attgttacga tttcctaaac ttcatgatgc catagttgaa gtggtgactt gtcttcttcg
     2881 taaaaggttg cctgttacaa atgaaatggt ccataactta gtggcaattg aactggctta
     2941 tatcaacaca aaacatccag actttgctga tgcttgtggg ctaatgaaca ataatataga
     3001 ggaacaaagg agaaacaggc tagccagaga attaccttca gctgtatcac gagacaaggt
     3061 tgcatctgga ggtggtgggg ttggagatgg tgttcaagaa ccaaccacag gcaactggag
     3121 aggaatgctg aaaacttcaa aagctgaaga gttattagca gaagaaaaat caaaacccat
     3181 tccaattatg ccagccagtc cacaaaaagg tcatgccgtg aacctgctag atgtgccagt
     3241 tcctgttgca cgaaaactat ctgctcggga acagcgagat tgtgaggtta ttgaacgact
     3301 cattaaatca tattttctca ttgtcagaaa gaatattcaa gacagtgtgc caaaggcagt
     3361 aatgcatttt ttggttaatc atgtgaaaga cactcttcag agtgagctag taggccagct
     3421 gtataaatca tccttattgg atgatcttct gacagaatct gaggacatgg cacagcgcag
     3481 gaaagaagca gctgatatgc taaaggcatt acaaggagcc agtcaaatta ttgctgaaat
     3541 ccgggagact catctttggt ggtctcatcc tcagttcgaa aaataagatc cggctgctaa
     3601 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3661 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     3721 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
     3781 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
     3841 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtcaa
     3901 acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3961 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4021 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4081 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4141 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4201 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4261 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4321 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     4381 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4441 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4501 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4561 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4621 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     4681 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4741 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4801 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4861 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4921 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4981 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5041 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5281 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5761 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5821 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5881 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     5941 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
     6001 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     6061 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6121 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6181 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6241 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6301 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
     6361 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
     6421 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
     6481 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
     6541 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
     6601 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
     6661 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
     6721 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
     6781 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
     6841 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
     6901 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
     6961 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
     7021 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
     7081 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
     7141 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
     7201 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
     7261 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
     7321 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     7381 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     7441 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     7501 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     7561 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     7621 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     7681 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7741 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     7801 gattgccctt cacc
//
