LOCUS       sequence-332911-        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-R465W
FEATURES             Location/Qualifiers
     source          1..8760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..80)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        65..394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      245..380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     307..326
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             461..1255
                     /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(515..534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1125..1144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1429..1550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1599..1615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1599..1615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1612..1634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1623..1639
                     /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(1647..1653,1654..1671,1672..1677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1692..1713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1830..1847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2001..2589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2081..2100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2760..3551,3552..3620))
                     /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     3383..3402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3621..3725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3800..3819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3991..4370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4371..4574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4525..4545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4619..4638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4619..4637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4742..4751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5501..5512
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7614..7748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8107..8124)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8113..8337
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8383..8760,1..51)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8470..8489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8680..8701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
       61 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      121 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      181 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      241 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      301 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      361 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
      421 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      481 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      541 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      601 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
      661 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      721 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      781 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      841 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      901 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      961 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1021 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1081 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1141 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1201 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1261 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     1321 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1381 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1441 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1501 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     1561 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     1621 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1681 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     1741 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2401 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3181 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3241 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3301 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3361 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3421 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3481 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3541 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3601 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3721 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     3781 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     3841 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     3901 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     3961 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4021 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4081 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4141 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4201 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4261 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4321 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4381 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4441 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4501 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     4561 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
     4621 atacgactca ctatagggag acccaagctg gctagcgttt aaacgggccc tctagactcg
     4681 agcggccgcc actgtgctgg atatctgcag aattcctgca gcccggggga tccaaggagc
     4741 cgccgccatg gagtatgatg ttcctgatta tgctcatatg ggcaaccgcg gcatggaaga
     4801 gctcatcccg ctggttaaca agctgcagga cgccttcagc tccatcggtc agagctgcca
     4861 cctggacctg ccgcagatcg ccgtggtggg cggccagagc gccggcaaga gttcggtgct
     4921 cgagaacttc gtgggccggg acttccttcc acgaggatca ggaattgtca cccggaggcc
     4981 tctcattctg cagctcatct tttccaaaac agaatatgcg gagtttttgc actgcaagtc
     5041 caaaaagttt acagactttg atgaagtccg gcaggagatc gaagcagaga ctgaccgggt
     5101 cacaggcacc aacaaaggca tctccccggt gcccatcaac cttcgggtct attcaccaca
     5161 cgtgttgaac ttgaccctca tcgacctccc aggcatcact aaggtgccag tgggggacca
     5221 gccaccagac atcgagtacc agatcaagga catgatcctg cagtttatca gccgggagag
     5281 cagcctcatt cttgccgtca cacctgccaa catggacttg gccaactcgg atgccctcaa
     5341 gctggccaag gaggtggacc cccaaggcct gcggaccatt ggtgtcatca ccaagctaga
     5401 cctgatggat gaaggcacag atgccaggga tgtcctggag aacaagctcc tgcccttgag
     5461 aagaggctac atcggcgtgg ttaaccgcag ccagaaagac atcgagggca gaaaggacat
     5521 ccgggcagct ctggcagctg agaggaaatt cttcctctcc cacccagcct accggcacat
     5581 ggctgaccgc atgggtaccc cacacttgca gaaaacactg aaccagcaac tgaccaacca
     5641 catccgggag tcactgccaa ccctgcgcag caagctgcag agccaactgc tgtccctgga
     5701 gaaggaagtg gaagagtaca agaatttccg gcctgacgac cccacacgca agaccaaagc
     5761 cctgctgcag atggtccagc agtttggagt ggactttgag aagcgaatcg aaggctctgg
     5821 agatcaagtc gacaccctgg agctgtctgg gggcgcccgc atcaatcgca tcttccacga
     5881 gcgctttccc tttgagctgg tgaagatgga gtttgacgag aaagacctcc gaagagagat
     5941 cagctacgcc attaagaaca tccacggagt cagaactggg ctcttcacgc cggatttggc
     6001 gttcgaggcc attgtgaaaa agcaggtggt caagctgaaa gagccctgtc tgaaatgcgt
     6061 ggacctggtt atccaggagc taatcagtac agttaggcaa tgcaccagca agctcagttc
     6121 ctacccccgc cttcgagagg agaccgaacg cattgtcacc acctacatct gggagcgaga
     6181 agggagaacc aaggaccaga tccttcttct gattgacatt gagcagtcct acatcaacac
     6241 aaaccatgaa gacttcattg gatttgccaa tgcccagcag aggagcacgc agctgaacaa
     6301 gaagagggcc atacctaacc agggggagat cttggtgatc cgcaggggct ggttgaccat
     6361 caacaacatc agcttgatga aaggcggttc caaggagtac tggttcgtgc tgactgctga
     6421 gtcgttgtct tggtacaagg atgaagagga aaaagaaaag aagtacatgc tgccactaga
     6481 caacctcaaa atacgggatg tggagaaggg cttcatgtcc aacaagcatg tgtttgccat
     6541 cttcaacaca gagcagagga acgtctacaa ggacctgcga cagatcgaac tggcttgtga
     6601 ctcccaggaa gatgtggaca gctggaaggc ttcgttcctg cgagctgggg tctacccaga
     6661 gaaggaccag gcagagaacg aggatggagc acaagagaac accttctcca tggacccgca
     6721 gctggagcgg caggtggaga ccatccgcaa cttggtggac tcctatgtgg ccatcatcaa
     6781 caagtccatc cgggacctca tgccaaagac catcatgcac ctcatgatca acaacacaaa
     6841 ggccttcatc caccatgagc tgctagccta cctgtactca tcagcagacc agagcagcct
     6901 catggaggag tcagctgagc aggctcagcg gcgggacgat atgctgcgca tgtaccatgc
     6961 actcaaggaa gcgctcaaca tcattgggga catcagcacc agcaccgtgt ccacgcctgt
     7021 gcccccacct gtcgatgata cttggctcca gaacaccagc agccacagcc ccactccaca
     7081 gcgcagacct gtgtccagtg tgcacccgcc gggccgccca cccgcagtgc ggggccccac
     7141 accggggcct cccctcattc ctatgcctgt gggggccaca tcctccttct cggcaccccc
     7201 catcccatcc cggcctggac cacagaatgt gtttgccaac aatgacccct tctctgcgcc
     7261 acctcagata ccgtctcggc cagcacggat tccccctggg atccctccag gagtgcccag
     7321 cagaagagcg cccgctgcgc ccagccggcc caccattatc cgcccagccg agccgtccct
     7381 gctcgactag gctgcagggg acagtgttct gggggggtcc tcgtgtacct acagtggtct
     7441 gggccctgct actgcccccc atgctgggac caggctcccc gtgggcaagc ctgccctctc
     7501 ctccctagcc ctagtgcaca tgataaggag ggagagggtg ggaaggaggc ccaaacaaac
     7561 agaacttggg ggctttgcac tttggggcga gggggagctc gtcagttcta gaggatccag
     7621 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     7681 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     7741 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     7801 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcctgcaa
     7861 gcctcgtcgt cctggccgga ccacgctatc tgtgcaaggt ccccggcccc ggacgcgcgc
     7921 tccatgagca gagcgcccgc cgccgaggcg aagactcggg cggcgccctg cccgtcccac
     7981 caggtcaaca ggcggtaacc ggcctcttca tcgggaatgc gcgcgacctt cagcatcgcc
     8041 ggcatgtccc cctggcggac gggaagtatc cagctcgacc aagcttaagt ttaaaccgct
     8101 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     8161 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     8221 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     8281 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     8341 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     8401 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     8461 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     8521 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     8581 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     8641 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     8701 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
//
