LOCUS       sequence-332910-        8760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of rat DNM2 fused to HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8760)
  AUTHORS   Puri C, Manni MM, Vicinanza M, Hilcenko C, Zhu Y, Runwal G, 
            Stamatakou E, Menzies FM, Mamchaoui K, Bitoun M, Rubinsztein DC
  TITLE     A DNM2 Centronuclear Myopathy Mutation Reveals a Link between 
            Recycling Endosome Scission and Autophagy.
  JOURNAL   Dev Cell. 2020 Apr 20;53(2):154-168.e6. doi: 
            10.1016/j.devcel.2020.03.018.
  PUBMED    32315611
REFERENCE   2  (bases 1 to 8760)
  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: 
            "10.1016/j.devcel.2020.03"; journalName: "Dev Cell"; date: 
            "2020-04-20- 20"; volume: "53"; issue: "2"; pages: "154-168.e6"
COMMENT     Sequence Label: pcDNA3-HA-DNM2-W525L
FEATURES             Location/Qualifiers
     source          1..8760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      752..761
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1511..1522
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3624..3758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4134)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4123..4347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4393..4821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4480..4499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4690..4711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4830..4850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4835..5164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5015..5150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5077..5096
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5231..6025
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5285..5304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5895..5914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6199..6320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6369..6385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6369..6385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6382..6404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6393..6409
                     /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)."
     promoter        complement(join(6417..6423,6424..6441,6442..6447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6462..6483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6600..6617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6771..7359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6851..6870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7530..8321,8322..8390))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8153..8172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8391..8495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8570..8589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagactcg agcggccgcc actgtgctgg atatctgcag
      721 aattcctgca gcccggggga tccaaggagc cgccgccatg gagtatgatg ttcctgatta
      781 tgctcatatg ggcaaccgcg gcatggaaga gctcatcccg ctggttaaca agctgcagga
      841 cgccttcagc tccatcggtc agagctgcca cctggacctg ccgcagatcg ccgtggtggg
      901 cggccagagc gccggcaaga gttcggtgct cgagaacttc gtgggccggg acttccttcc
      961 acgaggatca ggaattgtca cccggaggcc tctcattctg cagctcatct tttccaaaac
     1021 agaatatgcg gagtttttgc actgcaagtc caaaaagttt acagactttg atgaagtccg
     1081 gcaggagatc gaagcagaga ctgaccgggt cacaggcacc aacaaaggca tctccccggt
     1141 gcccatcaac cttcgggtct attcaccaca cgtgttgaac ttgaccctca tcgacctccc
     1201 aggcatcact aaggtgccag tgggggacca gccaccagac atcgagtacc agatcaagga
     1261 catgatcctg cagtttatca gccgggagag cagcctcatt cttgccgtca cacctgccaa
     1321 catggacttg gccaactcgg atgccctcaa gctggccaag gaggtggacc cccaaggcct
     1381 gcggaccatt ggtgtcatca ccaagctaga cctgatggat gaaggcacag atgccaggga
     1441 tgtcctggag aacaagctcc tgcccttgag aagaggctac atcggcgtgg ttaaccgcag
     1501 ccagaaagac atcgagggca gaaaggacat ccgggcagct ctggcagctg agaggaaatt
     1561 cttcctctcc cacccagcct accggcacat ggctgaccgc atgggtaccc cacacttgca
     1621 gaaaacactg aaccagcaac tgaccaacca catccgggag tcactgccaa ccctgcgcag
     1681 caagctgcag agccaactgc tgtccctgga gaaggaagtg gaagagtaca agaatttccg
     1741 gcctgacgac cccacacgca agaccaaagc cctgctgcag atggtccagc agtttggagt
     1801 ggactttgag aagcgaatcg aaggctctgg agatcaagtc gacaccctgg agctgtctgg
     1861 gggcgcccgc atcaatcgca tcttccacga gcgctttccc tttgagctgg tgaagatgga
     1921 gtttgacgag aaagacctcc gaagagagat cagctacgcc attaagaaca tccacggagt
     1981 cagaactggg ctcttcacgc cggatttggc gttcgaggcc attgtgaaaa agcaggtggt
     2041 caagctgaaa gagccctgtc tgaaatgcgt ggacctggtt atccaggagc taatcagtac
     2101 agttaggcaa tgcaccagca agctcagttc ctacccccgc cttcgagagg agaccgaacg
     2161 cattgtcacc acctacatca gggagcgaga agggagaacc aaggaccaga tccttcttct
     2221 gattgacatt gagcagtcct acatcaacac aaaccatgaa gacttcattg gatttgccaa
     2281 tgcccagcag aggagcacgc agctgaacaa gaagagggcc atacctaacc agggggagat
     2341 cttggtgatc cgcaggggct tgttgaccat caacaacatc agcttgatga aaggcggttc
     2401 caaggagtac tggttcgtgc tgactgctga gtcgttgtct tggtacaagg atgaagagga
     2461 aaaagaaaag aagtacatgc tgccactaga caacctcaaa atacgggatg tggagaaggg
     2521 cttcatgtcc aacaagcatg tgtttgccat cttcaacaca gagcagagga acgtctacaa
     2581 ggacctgcga cagatcgaac tggcttgtga ctcccaggaa gatgtggaca gctggaaggc
     2641 ttcgttcctg cgagctgggg tctacccaga gaaggaccag gcagagaacg aggatggagc
     2701 acaagagaac accttctcca tggacccgca gctggagcgg caggtggaga ccatccgcaa
     2761 cttggtggac tcctatgtgg ccatcatcaa caagtccatc cgggacctca tgccaaagac
     2821 catcatgcac ctcatgatca acaacacaaa ggccttcatc caccatgagc tgctagccta
     2881 cctgtactca tcagcagacc agagcagcct catggaggag tcagctgagc aggctcagcg
     2941 gcgggacgat atgctgcgca tgtaccatgc actcaaggaa gcgctcaaca tcattgggga
     3001 catcagcacc agcaccgtgt ccacgcctgt gcccccacct gtcgatgata cttggctcca
     3061 gaacaccagc agccacagcc ccactccaca gcgcagacct gtgtccagtg tgcacccgcc
     3121 gggccgccca cccgcagtgc ggggccccac accggggcct cccctcattc ctatgcctgt
     3181 gggggccaca tcctccttct cggcaccccc catcccatcc cggcctggac cacagaatgt
     3241 gtttgccaac aatgacccct tctctgcgcc acctcagata ccgtctcggc cagcacggat
     3301 tccccctggg atccctccag gagtgcccag cagaagagcg cccgctgcgc ccagccggcc
     3361 caccattatc cgcccagccg agccgtccct gctcgactag gctgcagggg acagtgttct
     3421 gggggggtcc tcgtgtacct acagtggtct gggccctgct actgcccccc atgctgggac
     3481 caggctcccc gtgggcaagc ctgccctctc ctccctagcc ctagtgcaca tgataaggag
     3541 ggagagggtg ggaaggaggc ccaaacaaac agaacttggg ggctttgcac tttggggcga
     3601 gggggagctc gtcagttcta gaggatccag acatgataag atacattgat gagtttggac
     3661 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3721 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3781 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     3841 aatgtggtat ggctgattat gatcctgcaa gcctcgtcgt cctggccgga ccacgctatc
     3901 tgtgcaaggt ccccggcccc ggacgcgcgc tccatgagca gagcgcccgc cgccgaggcg
     3961 aagactcggg cggcgccctg cccgtcccac caggtcaaca ggcggtaacc ggcctcttca
     4021 tcgggaatgc gcgcgacctt cagcatcgcc ggcatgtccc cctggcggac gggaagtatc
     4081 cagctcgacc aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
     4141 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4201 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4261 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4321 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4381 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4441 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4501 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4561 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4621 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4681 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4741 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4801 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4861 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4981 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5041 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5101 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5161 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     5221 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5281 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5341 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5401 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5461 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5521 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5581 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5641 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5701 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5761 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5821 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5881 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5941 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6001 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6061 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6121 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6181 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6241 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6301 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6361 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6421 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6481 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6541 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6601 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6661 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6721 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7381 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7501 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7561 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7621 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7681 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7741 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7801 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7861 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7921 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7981 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8041 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8101 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8161 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8221 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8281 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8341 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8401 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8461 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8521 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     8581 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8641 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     8701 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
