LOCUS       sequence-137704-        7245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mouse Drp1 isoform b (minus residues 
            514-602) with C-terminal His-tag and PreScission protease cleavage 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7245)
  AUTHORS   Liu R, Chan DC
  TITLE     The mitochondrial fission receptor Mff selectively recruits 
            oligomerized Drp1.
  JOURNAL   Mol Biol Cell. 2015 Dec 1;26(24):4466-77. doi: 
            10.1091/mbc.E15-08-0591. Epub 2015 Oct 7.
  PUBMED    26446846
REFERENCE   2  (bases 1 to 7245)
  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"; doi: 
            "10.1091/mbc.E15-08-0591"; journalName: "Mol Biol Cell"; date: 
            "2015-12-1- 1"; volume: "26"; issue: "24"; pages: "4466-77"
COMMENT     Sequence Label: pET21-Drp1(deltaIB)
FEATURES             Location/Qualifiers
     source          1..7245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(179..202)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             2049..2071
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2086..2110
                     /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(2110..2129)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2111..2129)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2213..2232)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2438..2515
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2516..3598
                     /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(2535..2554)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4407..4598
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4592..4614)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4700..4842
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4755..4774
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4857..4874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5028..5616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5108..5127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5787..6578,6579..6647))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6410..6429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6648..6752)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6779..7234)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6916..6937)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7128..7147
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgg
      181 gcccctggaa cagaacttcc agcggatccg cccaaagatg agtctctcgg atttcagcaa
      241 taatttgact ggctccttgt aatgccttca gcatatccgc tgcttctttt cttcgttggg
      301 ccatgtcctc ggattcagtc agaaggtcat ctaataagga tgacttatac agctgcccta
      361 ccagttcact ctgaagagta tctttcacat gattaaccaa aaaatgcatt actgcctttg
      421 ggacactgtc ttgaatattc tttctgacaa ttagaaaata tgatttgata agtctttcaa
      481 taacctcaca atctcgctgt tcctctatat tattgttcat tagcccacag gcatcagcaa
      541 agtcggggtg ttttgtgttg atatacgcta gctcaattgc cactaagtta tgcaccattt
      601 catttgtcac gggcaacctt ttacgaagaa gacaggtcac tacttcaact atggcatcgt
      661 gaagtttagg gaaccgtaac aattcctgtg tgctgtaatt gctacaatgc tgaatgatcc
      721 tctgcatctc ctcatggacc agctccacac accgtaggct gggctcttct agacgcttaa
      781 tctgacgttt gaccagtaac tcaaatgaaa cttcaggcac aaataaagca ggacgggggc
      841 cagtagcatt tctgatggca gtcaggatgt caatagtgtt aaggccacct agtgggtcca
      901 cagattctaa ggttcgcccg aaagtctcat ggaaaatata acaaatccta gcaccaccgc
      961 atagctcaga agtttcaatg tactttgcgg ttccttcaat cgtgttacaa tactctgtgg
     1021 caaatttggt aataagctgg agtaaagtag cacttttatc atccaccggt tcaccatagc
     1081 tatttagaag agactgatac tgagcagcta agacatttat tcttgttttc agctctggta
     1141 aacaatctct gatatgatgc ataagtaacc tattcagggt cctagcaaga tactttgttc
     1201 catttctgtt ggccagagat gggtacttct tttgaagaaa agcatactca tcacggattg
     1261 aatcagttac actcttctta ttgttaatat ccagttggct tctgttaact actccaatta
     1321 ttccaagctt gactggaata acccttccca tcaatacatc catggcatca gtacccgcat
     1381 ccatgagatc aagtttagtg attacagcta gagttctgcg cccatctgga tctacctctc
     1441 tggaaatctt gagtgcttct gatgttgcca tatctgtatt tgcagcagtg acggcgagga
     1501 taatggaatt gggattactg atgaaccgaa gaataagttc tctgatctga agctcgatgt
     1561 ccttgggctg atcgcctaca ggtactttgg tcattcccgg taaatccaca agtgtcaggt
     1621 tgacaacgtt gggcgagaaa accttgagat ggattggctc agggcttacc cccttattat
     1681 ttcctgaaat tctttcagtt tcattttcaa tttcttgtcg aatttcatca aaatctgtgt
     1741 aaagcttgtt tttggtgtga agaaatttac cccattcttc tgcttcaact ccattttctt
     1801 ctcctgttgt ttttctttta tcttctggtg aaacgtggac tagctgcaga atgagaggtc
     1861 tccgggtgac cacaccagtt cctctgggaa gaaggtccct gcccactagg ctttccagca
     1921 ctgagctctt cccactgctc tgcgttccca ctacgacgat ctgaggcagc tggatgatgt
     1981 ccgcacccac tgtgttgaag acgtcctgca gcttattgat gaccgggatc agcgcctcca
     2041 tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc
     2101 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac
     2161 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     2221 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     2281 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
     2341 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
     2401 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
     2461 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     2521 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     2581 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     2641 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     2701 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     2761 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     2821 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     2881 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     2941 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     3001 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     3061 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     3121 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     3181 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     3241 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     3301 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     3361 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     3421 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     3481 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     3541 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3601 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
     3661 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     3721 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     3781 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     3841 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     3901 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
     3961 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
     4021 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
     4081 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
     4141 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
     4201 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
     4261 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
     4321 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
     4381 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
     4441 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
     4501 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
     4561 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4621 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4681 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     4741 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt
     4801 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     4861 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4921 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4981 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5041 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5101 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5161 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5221 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5281 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5341 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5401 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5461 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5521 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5581 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5641 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5761 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5821 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5881 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5941 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6001 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6061 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     6121 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6181 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6241 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6301 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6361 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6421 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6481 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6541 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6601 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6661 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6721 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa
     6781 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     6841 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     6901 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     6961 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     7021 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     7081 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     7141 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     7201 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgcca
//
