LOCUS       sequence-379811-        8470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Mfn1, a mediator of mitochondrial fusion..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8470)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8470)
  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            "unpublished"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-Mfn1
FEATURES             Location/Qualifiers
     source          1..8470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1752..2207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1839..1858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2049..2070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2333..2355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2347..2364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2348..2364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2525..2904
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2905..3108
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3059..3079
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3151..3160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3169..3198
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    5438..6060
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5637..5656)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(6084..6104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6102..6399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6453..7247
                     /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(6507..6526)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7117..7136
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7899..8020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7936..7955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7990..8009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8036..8055)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8037..8055)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8069..8085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8069..8085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8082..8104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8093..8109
                     /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(8117..8123,8124..8141,8142..8147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8162..8183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8300..8317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1801 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2101 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2161 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     2221 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2281 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     2341 cgacgttgta aaacgacggc cagtgccaag ctgatctata cattgaatca atattggcaa
     2401 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     2461 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     2521 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     2581 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     2641 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2701 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2761 catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg taaatggccc
     2821 gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca gtacatctac
     2881 gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa tgggcgtgga
     2941 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     3001 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc cccgttgacg
     3061 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     3121 cgtcagaatt aattcagatc ggaattcgcc gccaccatgg gaggtaccga acaaaaactc
     3181 atctcagaag aggatctgtc tagtatggca gaacctgttt ctccactgaa gcactttgtg
     3241 ctggctaaga aggcgattac tgcaatcttt gaccagttac tggagtttgt tactgaagga
     3301 tcacattttg ttgaagcaac atataagaat ccggaacttg atcgaatagc cactgaagat
     3361 gatctggtag aaatgcaagg atataaagac aagctttcca tcattggtga ggtgctatct
     3421 cggagacaca tgaaggtggc attttttggc aggacaagca gtgggaagag ctctgttatc
     3481 aatgcaatgt tgtgggataa agttctccct agtgggattg gccatataac caattgcttc
     3541 ctaagtgttg aaggaactga tggagataaa gcctatctta tgacagaagg atcagatgaa
     3601 aaaaagagtg tgaagacagt taatcaactg gcccatgccc ttcacatgga caaagatttg
     3661 aaagctggct gtcttgtacg tgtgttttgg ccaaaagcaa aatgtgccct cttgagagat
     3721 gacctggtgt tagtagacag tccaggcaca gatgtcacta cagagctgga tagctggatt
     3781 gataagtttt gcctagatgc tgatgtcttt gttttggtcg caaactctga atcaacacta
     3841 atgaatacgg aaaaacactt ttttcacaag gtgaatgagc ggctttccaa gcctaatatt
     3901 ttcattctca ataatcgttg ggatgcctct gcatcagagc cagaatatat ggaagacgta
     3961 cgcagacagc acatggaaag atgcctgcat ttctcggtgg aggagctcaa agttgtaaat
     4021 gccttagaag cacagaatcg tatcttcttt gtttcagcaa aggaagttct tagtgctaga
     4081 aagcaaaaag cacaggggat gccagaaagt ggtgtggcac ttgctgaagg atttcatgca
     4141 agattacagg aatttcagaa ttttgaacaa atctttgagg agtgtatctc gcagtcagca
     4201 gtgaaaacaa agttcgaaca gcacactatc agagctaaac agatactagc tactgtgaaa
     4261 aacataatgg attcagtaaa cctggcagct gaagataaaa ggcattattc agtggaagag
     4321 agggaagacc aaattgatag actggacttt attcgaaacc agatgaacct tttaacactg
     4381 gatgttaaga aaaaaatcaa ggaggttacc gaggaggtgg caaacaaagt ttcatgtgca
     4441 atgacagatg aaatttgtcg actgtctgtt ttggttgatg aattttgttc agagtttcat
     4501 cctaatccag atgtattaaa aatatataaa agtgaattaa ataagcacat agaggatggt
     4561 atgggaagaa atttggctga tcgatgcacc gatgaagtaa acgccttagt gcttcagacc
     4621 cagcaagaaa ttattgaaaa tttgaagcca ttacttccag ctggtataca ggataaacta
     4681 catacactga tcccttgcaa gaaatttgat ctcagttata atctaaatta ccacaagtta
     4741 tgttcagatt ttcaagagga tattgtattt cgtttttccc tgggctggtc ttcccttgta
     4801 catcgatttt tgggccctag aaatgctcaa agggtgctcc taggattatc agagcctatc
     4861 tttcagctcc ctagatcttt agcttctact cccactgctc ctaccactcc agcaacgcca
     4921 gataatgcat cacaggaaga actcatgatt acattagtaa caggattggc gtccgttaca
     4981 tctagaactt ctatgggcat cattattgtt ggaggagtga tttggaaaac tataggctgg
     5041 aaactcctat ctgtttcatt aactatgtat ggagctttgt atctttatga aagactgagc
     5101 tggaccaccc atgccaagga gcgagccttt aaacagcagt ttgtaaacta tgcaactgaa
     5161 aaactgagga tgattgttag ctccacgagt gcaaactgca gtcaccaagt aaaacaacaa
     5221 atagctacca cttttgctcg cctgtgccaa caagttgata ttactcaaaa acagctggaa
     5281 gaagaaattg ctagattacc caaagaaata gatcagttgg agaaaataca aaacaattca
     5341 aagctcttaa gaaataaagc tgttcaactt gaaaatgagc tggagaattt tactaagcag
     5401 tttctacctt caagcaatga agaatcctaa ggatcccggg tggcatccct gtgacccctc
     5461 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     5521 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     5581 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     5641 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     5701 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     5761 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     5821 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     5881 acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca
     5941 ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag
     6001 ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt
     6061 gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     6181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggaattgatc agcttgggat
     6421 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     6481 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     6541 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     6601 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     6661 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6721 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6781 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6841 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6901 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6961 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     7021 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     7081 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     7141 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     7201 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     7261 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     7321 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     7381 ccagcgcggg gatctcatgc tggagttctt cgcccacccc gggctcgatc ccctcgcgag
     7441 ttggttcagc tgctgcctga ggctggacga cctcgcggag ttctaccggc agtgcaaatc
     7501 cgtcggcatc caggaaacca gcagcggcta tccgcgcatc catgcccccg aactgcagga
     7561 gtggggaggc acgatggccg ctttggtcga cccggacggg acgctcctgc gcctgataca
     7621 gaacgaattg cttgcaggca tctcatgagt gtgtcttccc gttttccgcc tgaggtcact
     7681 gcgtggatgg agcgctggcg cctgctgcgc gacggcgagc tgctcaccac ccactcgcca
     7741 agctggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg ccgatcataa
     7801 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     7861 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     7921 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7981 attctagttg tggtttgtcc aaactcatca atgtatctta tcatctggat caattcccta
     8041 tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     8101 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     8161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     8281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     8341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8461 tgctggcgtt
//
