LOCUS       sequence-379603-        7405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Mfn2 tagged with mitochondrion localization signal 
            (ActA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7405)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7405)
  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: 
            "unpublished"
COMMENT     Sequence Label: pcDNA3.1/Hygro-Mfn2-ActA
FEATURES             Location/Qualifiers
     source          1..7405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(130..718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(210..229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(889..1680,1681..1749))
                     /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     1512..1531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1750..1854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1929..1948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2120..2499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2500..2703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2654..2674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2748..2767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2748..2766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2802..2811
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2820..2849
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     RBS             3770..3778
                     /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)"
     primer_bind     complement(5298..5315)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5304..5528
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5574..6002
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5661..5680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5871..5892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6011..6031)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6016..6345
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6196..6331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6258..6277
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6393..6440
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6459..6833
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6963..7084
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7000..7019)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7054..7073
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7133..7149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7133..7149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7146..7168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7157..7173
                     /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(7181..7187,7188..7205,7206..7211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7226..7247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7364..7381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
       61 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1861 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1921 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     1981 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     2041 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     2101 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     2161 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2221 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2281 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2341 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2401 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2461 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2521 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2581 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2641 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2701 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     2761 tatagggaga cccaagctgg ctagcgttta aacttaagct tgccaccatg ggaggtaccg
     2821 aacaaaaact catctcagaa gaggatctgt ctagatccct gctcttctct cgatgcaact
     2881 ctatcgtcac agtcaagaaa aataagagac acatggctga ggtgaatgca tccccactta
     2941 agcactttgt cactgccaag aagaagatca atggcatttt tgagcagctg ggggcctaca
     3001 tccaggagag cgccaccttc cttgaagaca cgtacaggaa tgcagaactg gaccccgtta
     3061 ccacagaaga acaggttctg gacgtcaaag gttacctatc caaagtgaga ggcatcagtg
     3121 aggtgctggc tcggaggcac atgaaagtgg ctttttttgg ccggacgagc aatgggaaga
     3181 gcaccgtgat caatgccatg ctctgggaca aagttctgcc ctctgggatt ggccacacca
     3241 ccaattgctt cctgcgggta gagggcacag atggccatga ggcctttctc cttaccgagg
     3301 gctcagagga aaagaggagt gccaagactg tgaaccagct ggcccatgcc ctccaccagg
     3361 acaagcagct ccatgccggc agcctagtga gtgtgatgtg gcccaactct aagtgcccac
     3421 ttctgaagga tgacctcgtt ttgatggaca gccctggtat tgatgtcacc acagagctgg
     3481 acagctggat tgacaagttt tgtctggatg ctgatgtgtt tgtgctggtg gccaactcag
     3541 agtccaccct gatgcagacg gaaaagcact tcttccacaa ggtgagtgag cgtctctccc
     3601 ggccaaacat cttcatcctg aacaaccgct gggatgcatc tgcctcagag cccgagtaca
     3661 tggaggaggt gcggcggcag cacatggagc gttgtaccag cttcctggtg gatgagctgg
     3721 gcgtggtgga tcgatcccag gccggggacc gcatcttctt tgtgtctgct aaggaggtgc
     3781 tcaacgccag gattcagaaa gcccagggca tgcctgaagg agggggcgct ctcgcagaag
     3841 gctttcaagt gaggatgttt gagtttcaga attttgagag gagatttgag gagtgcatct
     3901 cccagtctgc agtgaagacc aagtttgagc agcacacggt ccgggccaag cagattgcag
     3961 aggcggttcg actcatcatg gactccctgc acatggcggc tcgggagcag caggtttact
     4021 gcgaggaaat gcgtgaagag cggcaagacc gactgaaatt tattgacaaa cagctggagc
     4081 tcttggctca agactataag ctgcgaatta agcagattac ggaggaagtg gagaggcagg
     4141 tgtcgactgc aatggccgag gagatcaggc gcctctctgt actggtggac gattaccaga
     4201 tggacttcca cccttctcca gtagtcctca aggtttataa gaatgagctg caccgccaca
     4261 tagaggaagg actgggtcga aacatgtctg accgctgctc cacggccatc accaactccc
     4321 tgcagaccat gcagcaggac atgatagatg gcttgaaacc cctccttcct gtgtctgtgc
     4381 ggagtcagat agacatgctg gtcccacgcc agtgcttctc cctcaactat gacctaaact
     4441 gtgacaagct gtgtgctgac ttccaggaag acattgagtt ccatttctct ctcggatgga
     4501 ccatgctggt gaataggttc ctgggcccca agaacagccg tcgggccttg atgggctaca
     4561 atgaccaggt ccagcgtccc atccctctga cgccagccaa ccccagcatg cccccactgc
     4621 cacagggctc gctcacccag gaggagttca tggtttccat ggttaccggc ctggcctcct
     4681 tgacatccag gacctccatg ggcattcttg ttgttggagg agtggtgtgg aaggcagtgg
     4741 gctggcggct cattgccctc tcctttgggc tctatggcct cctctacgtc tatgagcgtc
     4801 tgacctggac caccaaggcc aaggagaggg ccttcaagcg ccagtttgtg gagcatgcca
     4861 gcgagaagct gcagcttgtc atcagctaca ctggctccaa ctgcagccac caagtccagc
     4921 aggaactgtc tgggaccttt gctcatctgt gtcagcaagt tgacgtcacc cgggagaacc
     4981 tggagcagga aattgccgcc atgaacaaga aaattgaggt tcttgactca cttcagagca
     5041 aagcaaagct gctcaggaat aaagccggtt ggttggacag tgagctcaac atgttcacac
     5101 accagtacct gcagcccagc agactgattc tcgccatgct ggcaattggg gtgttctccc
     5161 tcggcgcctt cattaagatc attcagctcc ggaagaacaa ttagggatcc actagtccag
     5221 tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt
     5281 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     5341 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5401 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5461 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     5521 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     5581 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5641 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5701 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5761 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5821 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5881 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5941 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6001 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     6061 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     6121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     6181 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6241 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     6301 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     6361 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     6421 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg
     6481 ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg
     6541 ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc
     6601 tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg
     6661 tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg
     6721 acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc
     6781 tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc
     6841 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6901 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6961 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7021 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7081 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7321 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7381 tgcgctcggt cgttcggctg cggcg
//
