LOCUS       sequence-379667-        7288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the mitochondrial GTPase Mfn2. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7288)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7288)
  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            "unpublished"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-Mfn2
FEATURES             Location/Qualifiers
     source          1..7288
                     /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
     primer_bind     complement(1800..1819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1991..2370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2371..2574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2525..2545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2619..2638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2619..2637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3638..3646
                     /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(5052..5069)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5058..5282
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5328..5756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5415..5434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5625..5646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5765..5785)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5770..6099
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5950..6085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6012..6031
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6147..6194
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6213..6587
                     /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    6717..6838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6754..6773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6808..6827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6887..6903)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6887..6903)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6900..6922)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6911..6927
                     /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(6935..6941,6942..6959,6960..6965))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6980..7001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7118..7135)
                     /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 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     1801 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     1861 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     1921 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     1981 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2041 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2101 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2281 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2341 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2401 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2461 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2521 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2581 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2641 acccaagctg gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag
     2701 tgtggtggaa ttctgcagat atgtccctgc tcttctctcg atgcaactct atcgtcacag
     2761 tcaagaaaaa taagagacac atggctgagg tgaatgcatc cccacttaag cactttgtca
     2821 ctgccaagaa gaagatcaat ggcatttttg agcagctggg ggcctacatc caggagagcg
     2881 ccaccttcct tgaagacacg tacaggaatg cagaactgga ccccgttacc acagaagaac
     2941 aggttctgga cgtcaaaggt tacctatcca aagtgagagg catcagtgag gtgctggctc
     3001 ggaggcacat gaaagtggct ttttttggcc ggacgagcaa tgggaagagc accgtgatca
     3061 atgccatgct ctgggacaaa gttctgccct ctgggattgg ccacaccacc aattgcttcc
     3121 tgcgggtaga gggcacagat ggccatgagg cctttctcct taccgagggc tcagaggaaa
     3181 agaggagtgc caagactgtg aaccagctgg cccatgccct ccaccaggac aagcagctcc
     3241 atgccggcag cctagtgagt gtgatgtggc ccaactctaa gtgcccactt ctgaaggatg
     3301 acctcgtttt gatggacagc cctggtattg atgtcaccac agagctggac agctggattg
     3361 acaagttttg tctggatgct gatgtgtttg tgctggtggc caactcagag tccaccctga
     3421 tgcagacgga aaagcacttc ttccacaagg tgagtgagcg tctctcccgg ccaaacatct
     3481 tcatcctgaa caaccgctgg gatgcatctg cctcagagcc cgagtacatg gaggaggtgc
     3541 ggcggcagca catggagcgt tgtaccagct tcctggtgga tgagctgggc gtggtggatc
     3601 gatcccaggc cggggaccgc atcttctttg tgtctgctaa ggaggtgctc aacgccagga
     3661 ttcagaaagc ccagggcatg cctgaaggag ggggcgctct cgcagaaggc tttcaagtga
     3721 ggatgtttga gtttcagaat tttgagagga gatttgagga gtgcatctcc cagtctgcag
     3781 tgaagaccaa gtttgagcag cacacggtcc gggccaagca gattgcagag gcggttcgac
     3841 tcatcatgga ctccctgcac atggcggctc gggagcagca ggtttactgc gaggaaatgc
     3901 gtgaagagcg gcaagaccga ctgaaattta ttgacaaaca gctggagctc ttggctcaag
     3961 actataagct gcgaattaag cagattacgg aggaagtgga gaggcaggtg tcgactgcaa
     4021 tggccgagga gatcaggcgc ctctctgtac tggtggacga ttaccagatg gacttccacc
     4081 cttctccagt agtcctcaag gtttataaga atgagctgca ccgccacata gaggaaggac
     4141 tgggtcgaaa catgtctgac cgctgctcca cggccatcac caactccctg cagaccatgc
     4201 agcaggacat gatagatggc ttgaaacccc tccttcctgt gtctgtgcgg agtcagatag
     4261 acatgctggt cccacgccag tgcttctccc tcaactatga cctaaactgt gacaagctgt
     4321 gtgctgactt ccaggaagac attgagttcc atttctctct cggatggacc atgctggtga
     4381 ataggttcct gggccccaag aacagccgtc gggccttgat gggctacaat gaccaggtcc
     4441 agcgtcccat ccctctgacg ccagccaacc ccagcatgcc cccactgcca cagggctcgc
     4501 tcacccagga ggagttcatg gtttccatgg ttaccggcct ggcctccttg acatccagga
     4561 cctccatggg cattcttgtt gttggaggag tggtgtggaa ggcagtgggc tggcggctca
     4621 ttgccctctc ctttgggctc tatggcctcc tctacgtcta tgagcgtctg acctggacca
     4681 ccaaggccaa ggagagggcc ttcaagcgcc agtttgtgga gcatgccagc gagaagctgc
     4741 agcttgtcat cagctacact ggctccaact gcagccacca agtccagcag gaactgtctg
     4801 ggacctttgc tcatctgtgt cagcaagttg acgtcacccg ggagaacctg gagcaggaaa
     4861 ttgccgccat gaacaagaaa attgaggttc ttgactcact tcagagcaaa gcaaagctgc
     4921 tcaggaataa agccggttgg ttggacagtg agctcaacat gttcacacac cagtacctgc
     4981 agcccagcag atagatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac
     5041 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     5101 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     5161 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     5221 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     5281 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     5341 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5401 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5461 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5521 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5581 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5641 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5701 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     5761 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5821 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5881 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5941 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6001 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     6061 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     6121 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     6181 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     6241 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     6301 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     6361 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     6421 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     6481 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     6541 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     6601 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6661 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6721 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6781 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6841 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6901 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6961 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7021 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7081 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7141 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7261 accgtaaaaa ggccgcgttg ctggcgtt
//
