LOCUS       sequence-413164-        7730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminal HA tagged human Mfn2 S27D in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7730)
  AUTHORS   Leboucher GP, Tsai YC, Yang M, Shaw KC, Zhou M, Veenstra TD, 
            Glickman MH, Weissman AM
  TITLE     Stress-induced phosphorylation and proteasomal degradation of 
            mitofusin 2 facilitates mitochondrial fragmentation and apoptosis.
  JOURNAL   Mol Cell. 2012 Aug 24;47(4):547-57. doi: 
            10.1016/j.molcel.2012.05.041. Epub 2012 Jun 28.
  PUBMED    22748923
REFERENCE   2  (bases 1 to 7730)
  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.molcel.2012.05.041"; journalName: "Mol Cell"; date: 
            "2012-08-24- 24"; volume: "47"; issue: "4"; pages: "547-57"
COMMENT     Sequence Label: pcDNA3.1 Mfn2-HA S27D
FEATURES             Location/Qualifiers
     source          1..7730
                     /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"
     RBS             1596..1604
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2956..2982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2986..3012
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3049..3067)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3050..3067)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3087..3104)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3093..3317
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3363..3791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3450..3469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3660..3681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3800..3820)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3805..4134
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3985..4120
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4047..4066
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4201..4995
                     /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(4255..4274)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4865..4884
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5169..5290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5206..5225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5260..5279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5339..5355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5339..5355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5352..5374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5363..5379
                     /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(5387..5393,5394..5411,5412..5417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5432..5453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5570..5587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5741..6329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5821..5840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6500..7291,7292..7360))
                     /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     7123..7142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7361..7465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7540..7559)
                     /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 acccaagctt
      661 ggtaccgagc tcggatccat gtccctgctc ttctctcgat gcaactctat cgtcacagtc
      721 aagaaaaata agagacacat ggctgaggtg aatgcagacc cacttaagca ctttgtcact
      781 gccaagaaga agatcaatgg catttttgag cagctggggg cctacatcca ggagagcgcc
      841 accttccttg aagacacgta caggaatgca gaactggacc ccgttaccac agaagaacag
      901 gttctggacg tcaaaggtta cctatccaaa gtgagaggca tcagtgaggt gctggctcgg
      961 aggcacatga aagtggcttt ttttggccgg acgagcaatg ggaagagcac cgtgatcaat
     1021 gccatgctct gggacaaagt tctgccctct gggattggcc acaccaccaa ttgcttcctg
     1081 cgggtagagg gcacagatgg ccatgaggcc tttctcctta ccgagggctc agaggaaaag
     1141 aggagtgcca agactgtgaa ccagctggcc catgccctcc accaggacaa gcagctccat
     1201 gccggcagcc tagtgagtgt gatgtggccc aactctaagt gcccacttct gaaggatgac
     1261 ctcgttttga tggacagccc tggtattgat gtcaccacag agctggacag ctggattgac
     1321 aagttttgtc tggatgctga tgtgtttgtg ctggtggcca actcagagtc caccctgatg
     1381 cagacggaaa agcacttctt ccacaaggtg agtgagcgtc tctcccggcc aaacatcttc
     1441 atcctgaaca accgctggga tgcatctgcc tcagagcccg agtacatgga ggaggtgcgg
     1501 cggcagcaca tggagcgttg taccagcttc ctggtggatg agctgggcgt ggtggatcga
     1561 tcccaggccg gggaccgcat cttctttgtg tctgctaagg aggtgctcaa cgccaggatt
     1621 cagaaagccc agggcatgcc tgaaggaggg ggcgctctcg cagaaggctt tcaagtgagg
     1681 atgtttgagt ttcagaattt tgagaggaga tttgaggagt gcatctccca gtctgcagtg
     1741 aagaccaagt ttgagcagca cacggtccgg gccaagcaga ttgcagaggc ggttcgactc
     1801 atcatggact ccctgcacat ggcggctcgg gagcagcagg tttactgcga ggaaatgcgt
     1861 gaagagcggc aagaccgact gaaatttatt gacaaacagc tggagctctt ggctcaagac
     1921 tataagctgc gaattaagca gattacggag gaagtggaga ggcaggtgtc gactgcaatg
     1981 gccgaggaga tcaggcgcct ctctgtactg gtggacgatt accagatgga cttccaccct
     2041 tctccagtag tcctcaaggt ttataagaat gagctgcacc gccacataga ggaaggactg
     2101 ggtcgaaaca tgtctgaccg ctgctccacg gccatcacca actccctgca gaccatgcag
     2161 caggacatga tagatggctt gaaacccctc cttcctgtgt ctgtgcggag tcagatagac
     2221 atgctggtcc cacgccagtg cttctccctc aactatgacc taaactgtga caagctgtgt
     2281 gctgacttcc aggaagacat tgagttccat ttctctctcg gatggaccat gctggtgaat
     2341 aggttcctgg gccccaagaa cagccgtcgg gccttgatgg gctacaatga ccaggtccag
     2401 cgtcccatcc ctctgacgcc agccaacccc agcatgcccc cactgccaca gggctcgctc
     2461 acccaggagg agttcatggt ttccatggtt accggcctgg cctccttgac atccaggacc
     2521 tccatgggca ttcttgttgt tggaggagtg gtgtggaagg cagtgggctg gcggctcatt
     2581 gccctctcct ttgggctcta tggcctcctc tacgtctatg agcgtctgac ctggaccacc
     2641 aaggccaagg agagggcctt caagcgccag tttgtggagc atgccagcga gaagctgcag
     2701 cttgtcatca gctacactgg ctccaactgc agccaccaag tccagcagga actgtctggg
     2761 acctttgctc atctgtgtca gcaagttgac gtcacccggg agaacctgga gcaggaaatt
     2821 gccgccatga acaagaaaat tgaggttctt gactcacttc agagcaaagc aaagctgctc
     2881 aggaataaag ccggttggtt ggacagtgag ctcaacatgt tcacacacca gtacctgcag
     2941 cccagcagag aattctaccc atacgatgtt cctgactatg cgggctatcc ctatgacgtc
     3001 ccggactatg caggatccta actcgagcat gcatctagag ggccctattc tatagtgtca
     3061 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3121 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3181 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3241 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3301 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3361 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3421 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3481 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3541 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3601 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3661 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3721 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3781 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3901 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3961 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4021 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4081 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4141 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4201 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4261 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4321 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4381 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4441 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4501 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4561 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4621 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4681 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4741 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4801 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4861 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4921 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4981 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5041 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5101 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5161 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5221 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5281 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5341 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5401 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5461 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5521 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5581 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5641 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5701 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5761 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5821 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5881 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5941 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6001 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6061 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6121 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6181 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6241 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6301 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6361 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6421 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6481 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6541 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6601 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6661 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6721 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6781 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6841 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6901 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6961 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7021 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7081 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7141 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7201 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7261 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7321 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7381 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7441 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7501 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     7561 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7621 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7681 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
