LOCUS       sequence-452446-        7393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of SLC25A46 with R247A and R347A mutations in 
            mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7393)
  AUTHORS   Boopathy S, Luce BE, Lugo CM, Hakim P, McDonald J, Kim HL, Ponce J, 
            Ueberheide BM, Chao LH
  TITLE     Identification of SLC25A46 interaction interfaces with mitochondrial
            membrane fusogens Opa1 and Mfn2.
  JOURNAL   J Biol Chem. 2024 Aug 31:107740. doi: 10.1016/j.jbc.2024.107740.
  PUBMED    39222684
REFERENCE   2  (bases 1 to 7393)
  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: "J Biol 
            Chem. 2024 Aug 31:107740. doi: 10.1016/j.jbc.2024.107740."
COMMENT     Sequence Label: pcDNA3.1-SLC25A46[R257A,R347A]-P2A-Electra1 (p...
FEATURES             Location/Qualifiers
     source          1..7393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     228..248
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(257..685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(367..388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     579..598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    731..955
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     944..961
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1713..1769)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(3063..3082)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3064..3082)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3127..3330)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3156..3176)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3331..3710
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3882..3901
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3976..4080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4081..4149,4150..4941)
                     /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     complement(4299..4318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5112..5700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5601..5620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5854..5871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5988..6009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6024..6029,6030..6047,6048..6054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6062..6078
                     /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)."
     primer_bind     6067..6089
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6086..6102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6086..6102
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6151..6272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6162..6181)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6216..6235
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6446..7240)
                     /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(6557..6576)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7167..7186
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7307..7393,1..243))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7321..7393,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(join(7375..7393,1))
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      181 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      241 cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      301 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
      361 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
      421 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
      481 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
      541 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
      601 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
      661 cttaatgcgc cgctacaggg cgcgtgggga taccccctag agccccagct ggttctttcc
      721 gcctcagaag ccatagagcc caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc
      781 tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct actcagacaa
      841 tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc
      901 aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag
      961 gctgatcagc gggtttaaac gggccctcta gattacttgt acagctcgtc catgccacca
     1021 ccaaggttgc tgtatttggc aactgccact tctcttagca ctacgtaggt ttcattgtca
     1081 ctctcttcga ccctttccag gcggtaatca acggcatgga caccgggcat cttgatgtcc
     1141 ccgacggtct tttttgacct gtaagttgtc acgatgttgg cgtgcagatg gccatcacca
     1201 tcaactttca gtgcgatgtc ggtatatcct ctcagaccac catttgctgg atacatcatc
     1261 tctgtgaaag gctcccaacc ctcggtcttc ttctgcatca cgggaccatt ggagggaaag
     1321 tttacccctc tgaccttgac gttgtagacg agccccccat cctcaaggct ggtgtcctgc
     1381 gtgacggtga cgactccacc atcttcgtat ctcgtaactc tttcccaagt aaagccctca
     1441 ggaaaggact gtttgaagaa atcagggatg tcggccgggt acttgataaa ggtcttgctg
     1501 ccgaacagga acgacgtggc aagaatgtca aaggcaaatg gcaggggtcc tccctcgatg
     1561 actttgatcc tcatggtttg cactccctcg tacggtctgc cttctccttc acctgtgcat
     1621 ttgaattggt ggccgttgac cgaaccttcc atgaccacct tcatacgcat attttcctcg
     1681 atcagctctt cgcccttaga caccatgaat tcaggtccag ggttctcctc cacgtctcca
     1741 gcctgcttca gcaggctgaa gttagtagct ccgcttccgg atccgatatt gttctgcagc
     1801 agtgtggagt agatgatctt ggtgatctgc agcacggcgg cgtgcagggt atactggatg
     1861 atcacggcgc caaagccctt gtaaaagccg aacacgccct cctcctgtct gatggtgttg
     1921 atacaatccc gcatgccctc atactgtgta ttgattggca gcacctcgta gcccagatcg
     1981 gtgttgtcga tgattgtggc ggtgccctgg atgtgcagcc tgtgcagcac tgtctccaga
     2041 ggatacagga tcacatcgga gcacaggctg gcggcaaaat tggcgatcag ctcggggaaa
     2101 taggcgtcca gcatagactg cactggagat gtggactcgg ccaggtggga gttgtaggtc
     2161 ttcctcttca ggatcagcag cacaaacttc tggatcacag aggagatgat gtagtgcagc
     2221 acgccgtgca gcacggtagg gaagatcagg ctcagcaggg gcagcagggc cttagagtgt
     2281 ggcacgccca tgccgatcac tctgccgatg ccctccttca cgcactccag gatgcctgta
     2341 ttgtcccgga tgatctcgct ctgcacggtc tcgatcaggc tggcagaata gaagggcatg
     2401 gccaccacgt aggtcaggga cttcagcagc aggtgctcgc cgatctgctt tgggctccac
     2461 ttgtgcagca cctcgcgtgg cagaggtgta aactcggaga tgatgccctc tgctcccagt
     2521 gtcactccct gcacgatgaa ggtgctgccc attcccttcc acagggccct tggtccctgg
     2581 gtcttattaa aagagtacat gatgttgatc actgtgaaag gggtcaggtg atagtgctgg
     2641 gcgtggtaat tcacttgaca ctggcgcctc agcacgatgc aagggtgggc cagcacgttc
     2701 tctgtaaaca gggatgccag tccgattcca aatcctgcga acctattcag ctgctcgcta
     2761 gactgtccct gcacagatcc tcctcctccg gagctgaaag gctcctctgt ggggccctca
     2821 tatggggtgc ttgtggtagg cacgccgtag ggtggagact tctcgcccca gtgcaggttt
     2881 ctggagccgg ggatatcagg tggtgtggtc acccagtgtc ccaggtcgct tccggtggaa
     2941 aagctccgtg ctggaaatgc tcctccgaat ccctgctcgt ctcttgctcc tcccctgtat
     3001 cccagtccat cgaagccgtc aggtctccgg gggtgcatgg tggcgctagc cagcttgggt
     3061 ctccctatag tgagtcgtat taatttcgat aagccagtaa gcagtgggtt ctctagttag
     3121 ccagagagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     3181 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     3241 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     3301 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     3361 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     3421 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3481 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3541 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3601 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3661 taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata
     3721 tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca
     3781 attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac
     3841 ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt
     3901 actgagagtg caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact
     3961 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4021 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4081 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4141 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4201 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4261 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4321 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4381 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4441 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4501 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4561 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4621 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4681 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4741 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4801 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4861 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4921 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4981 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5041 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5221 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5461 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5641 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5761 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5821 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5881 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5941 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6001 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6061 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     6121 tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt ggacaaacca
     6181 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     6241 ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc cagcatgaga
     6301 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     6361 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     6421 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     6481 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     6541 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     6601 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     6661 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     6721 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     6781 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     6841 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     6901 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     6961 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     7021 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     7081 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     7141 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     7201 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     7261 tctcttgatc agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta
     7321 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg
     7381 cctctgcata aat
//
