LOCUS       sequence-343850-        8054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses S. pombe Drp1 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8054)
  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 8054)
  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)-yDrp1-StrepII
FEATURES             Location/Qualifiers
     source          1..8054
                     /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="ENLYFQM"
     CDS             5228..5251
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(5304..5322)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5318..5365
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5540..5545,5546..5562,5563..5568))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5595..5613)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5681..5785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5786..5854,5855..6646)
                     /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(6004..6023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6817..7405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7306..7325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7559..7576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7591..7730
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7656..7675)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7816..7838
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7832..8023)
                     /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 gtcgcccaac 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 ccaaatggaa caacttattc cattggtgaa tcaacttcaa gaccttgtct acaacacaat
     2941 tgggtcagat tttttggatt taccatccat agtggtcgtt gggtcccaat cttgtggaaa
     3001 atcttctgta ttggaaaata ttgttggtaa agatttttta cctcgtggta ccggaattgt
     3061 cactagaaga cccttgatac tacaactcat caacttgaag cgcaagacca aaaataacca
     3121 tgatgaggaa agtacttctg acaacaattc agaagaaaca agtgctgctg gggaaacagg
     3181 cagtcttgaa ggaattgaag aagattctga tgaaattgaa gattatgctg aatttttgca
     3241 tatccccgat accaaattta ccgacatgaa caaagttcgc gcagagattg agaatgagac
     3301 tctacgggtt gctggtgcaa acaaaggaat taacaaattg ccaatcaatc taaaaattta
     3361 ttctactcgc gttttgaact taactttaat tgatttgcct gggcttacaa aaattcccgt
     3421 tggagatcag cctactgata ttgaggctca aacacgctct cttattatgg aatacatcag
     3481 ccgtcccaat tccattatac ttgctgtatc tcctgccaac tttgacattg tcaacagtga
     3541 gggcctaaaa ctagcgaggt ctgttgatcc caagggtaag cgcactattg gtgttcttac
     3601 caagcttgac cttatggatc aaggtaccaa tgcaatggac atattaagcg gtcgcgttta
     3661 tccccttaag ctgggtttcg tagcaactgt taaccgttcc cagtctgata ttgtatctca
     3721 caaatccatg agagatgctt tgcaatctga gcgctctttt tttgaacatc atccggcata
     3781 tcgtaccatt aaagatcgtt gtggaacacc ctatcttgcc aagacattga gtaatctatt
     3841 ggtatcacat atccgtgagc gccttcccga tatcaaagcc cgccttagca ctctaatatc
     3901 tcaaactcaa caacaactaa acaattacgg tgactttaaa ctttctgatc agagtcaacg
     3961 tggaattatt ttattgcagg cgatgaatcg ctttgccaac acgttcattg ctagtataga
     4021 tggaaattct tcaaatattc ccacaaaaga actttcaggt ggtgctcgtc tttactccat
     4081 tttcaataat gtatttacta ctgcattgaa ttcaatcgat cctttacaaa acttatcgac
     4141 agtcgacata cgtacagcta tattgaatag cacaggctct agggctacct tgttcctttc
     4201 ggaaatggct ttcgatatat tggtgaagcc gcaattaaac cttttagcgg caccctgtca
     4261 tcaatgtgtc gagcttgttt acgaagaatt aatgaaaata tgtcattatt ctggtgactc
     4321 agacatatcg cattttccca agttgcaaac tgctctcgta gaaaccgtta gcgatttact
     4381 acgagaaaat ttaactccta cttattcttt tgtagagagc ttgattgcta ttcaaagtgc
     4441 atatattaat acgaatcatc cggatttctt aggggtgcag ggtgcaatgg ctgtagtatt
     4501 gtcaagaaaa gaacagaatc gcttaatgct ttcccaagaa aatgatgaac ccatttcttc
     4561 tgctttggac actgtgaagc ctgacggaat tgagttgtat tcttcggatc ccgacacttc
     4621 tgttaagagt attaccaata aagcgacgaa cgagattaca actttgaaga gcgatgattc
     4681 agcaaaaatg cagcctttgg atgttttggc ttctaaacgg tataataatg cctttagcac
     4741 tgaaactgcg gaaagaaaaa catttcttag ctatgtcttt ggtgccaata atgcgactag
     4801 gaaagcaatg agcattgata agtcttctag ctatccttta aatgattctc tttctggagg
     4861 agatacaaat cacaaaaaca accatccttt aaagatgaca gatttatcaa atgaagttga
     4921 aacaatggcg ttggaggaca tgagtgaaag ggaagaagtt gaagttgatc ttattaaaga
     4981 gctaattact tcttatttca acttaactag gaagattata attgatcagg taccaaaggt
     5041 aattatgcat ttgctcgtga acgcctctaa agatgccatt caaaatcgtt tggttagtaa
     5101 attataccga gaggatttct ttgacaccct ccttattgaa gatgagaacg tcaaaagtga
     5161 aagagaaaaa tgtgagcgtc ttttatctgt ttacaaccaa gcaaacaaaa ttatttcaac
     5221 ggttttttgg tctcatcctc agttcgaaaa ataagatccg gctgctaaca aagcccgaaa
     5281 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5341 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag
     5401 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg
     5461 aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt
     5521 aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaattc
     5581 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5641 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5701 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5761 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5821 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5881 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5941 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6001 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     6061 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6121 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6181 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6241 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6301 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     6361 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6421 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6481 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6541 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6601 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6661 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6721 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6781 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6841 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6901 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6961 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7021 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7081 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7141 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7201 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7261 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7321 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7381 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7441 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7501 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7561 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     7621 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7681 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     7741 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     7801 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     7861 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     7921 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     7981 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     8041 ggggatttct gttc
//
