LOCUS       sequence-345882-        8504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Drp1 Isoform 3 with EGFP at C-terminus in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8504)
  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 8504)
  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: Drp1-EGFP-StrepII
FEATURES             Location/Qualifiers
     source          1..8504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(479..1561)
                     /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     1523..1542
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1562..1639)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1845..1864
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1948..1967
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1948..1966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1967..1991
                     /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             2006..2028
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(4162..4164,4165..4875)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4204..4225)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4465..4484)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4812..4833
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4876..4899
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4952..4970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4966..5013
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5188..5193,5194..5210,5211..5216))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5243..5261)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5329..5433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5434..5502,5503..6294)
                     /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(5652..5671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6465..7053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6954..6973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7207..7224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7239..7378
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7304..7323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7464..7486
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7480..7671)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
       61 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
      121 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
      181 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
      241 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
      301 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
      361 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
      421 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
      481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      541 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
      601 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
      661 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
      721 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
      781 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
      841 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
      901 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
      961 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     1021 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     1081 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     1141 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     1201 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     1261 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     1321 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     1381 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     1441 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     1501 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     1561 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     1621 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     1681 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     1741 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     1801 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     1861 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     1921 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     1981 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc
     2041 agcagcggct cgatggaggc gctaattcct gtcataaaca agctccagga cgtcttcaac
     2101 acggtgggcg ccgacatcat ccagctgcct caaatcgtcg tagtgggaac gcagagcagc
     2161 ggaaagagct cagtgctaga aagcctggtg gggagggacc tgcttcccag aggtactgga
     2221 attgtcaccc ggagacctct cattctgcaa ctggtccatg tttcacaaga agataaacgg
     2281 aaaacaacag gagaagaaaa tggggtggaa gcagaagaat ggggtaaatt tcttcacacc
     2341 aaaaataagc tttacacgga ttttgatgaa attcgacaag aaattgaaaa tgaaacagaa
     2401 agaatttcag gaaataataa gggagtaagc cctgaaccaa ttcatcttaa gattttttca
     2461 cccaacgttg tcaatttgac acttgtggat ttgccaggaa tgaccaaggt gcctgtaggt
     2521 gatcaaccta aggatattga gcttcaaatc agagagctca ttcttcggtt catcagtaat
     2581 cctaattcca ttatcctcgc tgtcactgct gctaatacag atatggcaac atcagaggca
     2641 cttaaaattt caagagaggt agatccagat ggtcgcagaa ccctagctgt aatcactaaa
     2701 cttgatctca tggatgcggg tactgatgcc atggatgtat tgatgggaag ggttattcca
     2761 gtcaaacttg gaataattgg agtagttaac aggagccagc tagatattaa caacaagaag
     2821 agtgtaactg attcaatccg tgatgagtat gcttttcttc aaaagaaata tccatctctg
     2881 gccaatagaa atggaacaaa gtatcttgct aggactctaa acaggttact gatgcatcac
     2941 atcagagatt gtttaccaga gttgaaaaca agaataaatg ttctagctgc tcagtatcag
     3001 tctcttctaa atagctacgg tgaacccgtg gatgataaaa gtgctacttt actccaactt
     3061 attaccaaat ttgccacaga atattgtaac actattgaag gaactgcaaa atatattgaa
     3121 acttcggagc tatgcggtgg tgctagaatt tgttatattt tccatgagac ttttgggcga
     3181 accttagaat ctgttgatcc acttggtggc cttaacacta ttgacatttt gactgccatt
     3241 agaaatgcta ctggtcctcg tcctgcttta tttgtgcctg aggtttcatt tgagttactg
     3301 gtgaagcggc aaatcaaacg tctagaagag cccagcctcc gctgtgtgga actggttcat
     3361 gaggaaatgc aaaggatcat tcagcactgt agcaattaca gtacacagga attgttacga
     3421 tttcctaaac ttcatgatgc catagttgaa gtggtgactt gtcttcttcg taaaaggttg
     3481 cctgttacaa atgaaatggt ccataactta gtggcaattg aactggctta tatcaacaca
     3541 aaacatccag actttgctga tgcttgtggg ctaatgaaca ataatataga ggaacaaagg
     3601 agaaacaggc tagccagaga attaccttca gctgtatcac gagacaaggt tgcatctgga
     3661 ggtggtgggg ttggagatgg tgttcaagaa ccaaccacag gcaactggag aggaatgctg
     3721 aaaacttcaa aagctgaaga gttattagca gaagaaaaat caaaacccat tccaattatg
     3781 ccagccagtc cacaaaaagg tcatgccgtg aacctgctag atgtgccagt tcctgttgca
     3841 cgaaaactat ctgctcggga acagcgagat tgtgaggtta ttgaacgact cattaaatca
     3901 tattttctca ttgtcagaaa gaatattcaa gacagtgtgc caaaggcagt aatgcatttt
     3961 ttggttaatc atgtgaaaga cactcttcag agtgagctag taggccagct gtataaatca
     4021 tccttattgg atgatcttct gacagaatct gaggacatgg cacagcgcag gaaagaagca
     4081 gctgatatgc taaaggcatt acaaggagcc agtcaaatta ttgctgaaat ccgggagact
     4141 catctttggc tcgaggatcc ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4201 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     4261 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4321 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     4381 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4441 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4501 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     4561 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     4621 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     4681 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4741 ctgctgcccg acaaccacta cctgagcacc cagtccaagt tgagcaaaga ccccaacgag
     4801 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4861 gacgagctgt acaagtggtc tcatcctcag ttcgaaaaat aagatccggc tgctaacaaa
     4921 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     4981 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat
     5041 cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga
     5101 cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt
     5161 agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat
     5221 gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5281 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5341 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5401 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5461 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5521 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5581 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5641 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5701 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5761 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5821 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5881 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5941 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     6001 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6061 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6121 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6181 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6241 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6301 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6361 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6421 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6481 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6541 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6601 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6661 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6721 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6781 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6841 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6901 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6961 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7021 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7081 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     7141 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     7201 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     7261 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     7321 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     7381 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     7441 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     7501 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     7561 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     7621 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     7681 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     7741 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     7801 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     7861 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     7921 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     7981 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     8041 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     8101 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     8161 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     8221 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     8281 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     8341 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     8401 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     8461 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctg
//
