LOCUS       sequence-137706-        7512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mouse Drp1 isoform b (699amino acids) 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 7512)
  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 7512)
  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
FEATURES             Location/Qualifiers
     source          1..7512
                     /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             2316..2338
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2353..2377
                     /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(2377..2396)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2378..2396)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2480..2499)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2705..2782
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2783..3865
                     /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(2802..2821)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4674..4865
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4859..4881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4967..5109
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5022..5041
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5124..5141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5295..5883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5375..5394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6054..6845,6846..6914))
                     /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     6677..6696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6915..7019)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7046..7501)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7183..7204)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7395..7414
                     /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 tctcgggcag acagttttct tgcaactgga actggcacat
      541 ctagcaaatt gacagcatgg cctttctgtg gacttgctgg cataattgga attggttttg
      601 atttttcttc agcaagtaat tcttcagctt ttgaagtttt cagcattcct ctccagttgc
      661 ctgttgttgg ttcctgacca ccgtctccaa tcccaccacc tgcagatggg accttgtccc
      721 gtgatccagc tgaaggcagc tctcttgcta gcctgtttct tctttgttcc tctatattat
      781 tgttcattag cccacaggca tcagcaaagt cggggtgttt tgtgttgata tacgctagct
      841 caattgccac taagttatgc accatttcat ttgtcacggg caacctttta cgaagaagac
      901 aggtcactac ttcaactatg gcatcgtgaa gtttagggaa ccgtaacaat tcctgtgtgc
      961 tgtaattgct acaatgctga atgatcctct gcatctcctc atggaccagc tccacacacc
     1021 gtaggctggg ctcttctaga cgcttaatct gacgtttgac cagtaactca aatgaaactt
     1081 caggcacaaa taaagcagga cgggggccag tagcatttct gatggcagtc aggatgtcaa
     1141 tagtgttaag gccacctagt gggtccacag attctaaggt tcgcccgaaa gtctcatgga
     1201 aaatataaca aatcctagca ccaccgcata gctcagaagt ttcaatgtac tttgcggttc
     1261 cttcaatcgt gttacaatac tctgtggcaa atttggtaat aagctggagt aaagtagcac
     1321 ttttatcatc caccggttca ccatagctat ttagaagaga ctgatactga gcagctaaga
     1381 catttattct tgttttcagc tctggtaaac aatctctgat atgatgcata agtaacctat
     1441 tcagggtcct agcaagatac tttgttccat ttctgttggc cagagatggg tacttctttt
     1501 gaagaaaagc atactcatca cggattgaat cagttacact cttcttattg ttaatatcca
     1561 gttggcttct gttaactact ccaattattc caagcttgac tggaataacc cttcccatca
     1621 atacatccat ggcatcagta cccgcatcca tgagatcaag tttagtgatt acagctagag
     1681 ttctgcgccc atctggatct acctctctgg aaatcttgag tgcttctgat gttgccatat
     1741 ctgtatttgc agcagtgacg gcgaggataa tggaattggg attactgatg aaccgaagaa
     1801 taagttctct gatctgaagc tcgatgtcct tgggctgatc gcctacaggt actttggtca
     1861 ttcccggtaa atccacaagt gtcaggttga caacgttggg cgagaaaacc ttgagatgga
     1921 ttggctcagg gcttaccccc ttattatttc ctgaaattct ttcagtttca ttttcaattt
     1981 cttgtcgaat ttcatcaaaa tctgtgtaaa gcttgttttt ggtgtgaaga aatttacccc
     2041 attcttctgc ttcaactcca ttttcttctc ctgttgtttt tcttttatct tctggtgaaa
     2101 cgtggactag ctgcagaatg agaggtctcc gggtgaccac accagttcct ctgggaagaa
     2161 ggtccctgcc cactaggctt tccagcactg agctcttccc actgctctgc gttcccacta
     2221 cgacgatctg aggcagctgg atgatgtccg cacccactgt gttgaagacg tcctgcagct
     2281 tattgatgac cgggatcagc gcctccatat gtatatctcc ttcttaaagt taaacaaaat
     2341 tatttctaga ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt
     2401 cgcgggatcg agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     2461 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     2521 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     2581 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     2641 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat
     2701 cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga
     2761 gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc
     2821 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa
     2881 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc
     2941 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct
     3001 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
     3061 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
     3121 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
     3181 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
     3241 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
     3301 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
     3361 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
     3421 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
     3481 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
     3541 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata
     3601 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
     3661 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
     3721 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
     3781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3841 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
     3901 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     3961 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     4021 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     4081 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     4141 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     4201 cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     4261 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     4321 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     4381 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     4441 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     4501 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     4561 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     4621 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     4681 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     4741 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg
     4801 accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     4861 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4921 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     4981 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     5041 actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5101 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5161 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5221 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5281 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5341 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5401 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5461 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5521 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5581 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5641 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5701 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5761 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5821 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5881 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5941 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6001 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6061 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6121 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6181 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6241 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6301 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6361 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6421 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6481 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6541 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6601 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6661 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6721 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6781 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6841 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6901 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6961 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7021 acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt aaaattcgcg
     7081 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     7141 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     7201 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     7261 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     7321 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     7381 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     7441 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     7501 tcccattcgc ca
//
