LOCUS       sequence-463588-        7393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of SLC25A46 with G249D mutation 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[G249D]-P2A-Electra1 (pAH10)
FEATURES             Location/Qualifiers
     source          1..7393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2176..2232
                     /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(2984..3001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2990..3214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3260..3688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3347..3366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3557..3578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3697..3717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3702..4031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3882..4017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3944..3963
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4098..4892
                     /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(4152..4171)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4762..4781
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5066..5187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5103..5122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5157..5176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5236..5252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5236..5252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5249..5271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5260..5276
                     /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(5284..5290,5291..5308,5309..5314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5329..5350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5467..5484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5638..6226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5718..5737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6397..7188,7189..7257))
                     /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     7020..7039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7258..7362)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgc acccccggag acctgacggc ttcgatggac tgggatacag gggaggagca
      961 agagacgagc agggattcgg aggagcattt ccagcacgga gcttttccac cggaagcgac
     1021 ctgggacact gggtgaccac accacctgat atccccggct ccagaaacct gcactggggc
     1081 gagaagtctc caccctacgg cgtgcctacc acaagcaccc catatgaggg ccccacagag
     1141 gagcctttca gctccggagg aggaggatct gtgcagggac agtctagcga gcagctgaat
     1201 aggttcgcag gatttggaat cggactggca tccctgttta cagagaacgt gctggcccac
     1261 ccttgcatcg tgctgaggcg ccagtgtcaa gtgaattacc acgcccagca ctatcacctg
     1321 acccctttca cagtgatcaa catcatgtac tcttttaata agacccaggg accaagggcc
     1381 ctgtggaagg gaatgggcag caccttcatc gtgcagggag tgacactggg agcagagggc
     1441 atcatctccg agtttacacc tctgccacgc gaggtgctgc acaagtggag cccaaagcag
     1501 atcggcgagc acctgctgct gaagtccctg acctacgtgg tggccatgcc cttctattct
     1561 gccagcctga tcgagaccgt gcagagcgag atcatccggg acaatacagg catcctggag
     1621 tgcgtgaagg agggcatcgg cagagtgatc gacatgggcg tgccacactc taagcgcctg
     1681 ctgcccctgc tgagcctgat cttccctacc gtgctgcacg gcgtgctgca ctacatcatc
     1741 tcctctgtga tccagaagtt tgtgctgctg atcctgaaga ggaagaccta caactcccac
     1801 ctggccgagt ccacatctcc agtgcagtct atgctggacg cctatttccc cgagctgatc
     1861 gccaattttg ccgccagcct gtgctccgat gtgatcctgt atcctctgga gacagtgctg
     1921 cacaggctgc acatccaggg cacccgcaca atcatcgaca acaccgatct gggctacgag
     1981 gtgctgccaa tcaatacaca gtatgagggc atgcgggatt gtatcaacac catcagacag
     2041 gaggagggcg tgttcggctt ttacaagggc tttggcgccg tgatcatcca gtataccctg
     2101 cacgccgccg tgctgcagat caccaagatc atctactcca cactgctgca gaacaatatc
     2161 ggatccggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     2221 aaccctggac ctgaattcat ggtgtctaag ggcgaagagc tgatcgagga aaatatgcgt
     2281 atgaaggtgg tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga
     2341 gaaggcagac cgtacgaggg agtgcaaacc atgaggatca aagtcatcga gggaggaccc
     2401 ctgccatttg cctttgacat tcttgccacg tcgttcctgt tcggcagcaa gacctttatc
     2461 aagtacccgg ccgacatccc tgatttcttc aaacagtcct ttcctgaggg ctttacttgg
     2521 gaaagagtta cgagatacga agatggtgga gtcgtcaccg tcacgcagga caccagcctt
     2581 gaggatgggg ggctcgtcta caacgtcaag gtcagagggg taaactttcc ctccaatggt
     2641 cccgtgatgc agaagaagac cgagggttgg gagcctttca cagagatgat gtatccagca
     2701 aatggtggtc tgagaggata taccgacatc gcactgaaag ttgatggtga tggccatctg
     2761 cacgccaaca tcgtgacaac ttacaggtca aaaaagaccg tcggggacat caagatgccc
     2821 ggtgtccatg ccgttgatta ccgcctggaa agggtcgaag agagtgacaa tgaaacctac
     2881 gtagtgctaa gagaagtggc agttgccaaa tacagcaacc ttggtggtgg catggacgag
     2941 ctgtacaagt aatctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3001 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3061 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3121 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3181 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3241 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3301 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3361 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3421 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3481 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3541 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3601 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3661 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3721 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3961 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4021 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4081 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4141 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4201 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4261 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4321 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4381 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4441 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4501 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4561 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4621 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4681 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4741 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4801 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4861 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4921 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4981 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5041 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5101 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5161 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5221 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5281 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5341 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5401 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5461 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5521 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5581 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5641 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5701 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5761 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5821 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5881 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5941 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6001 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6061 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6361 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6421 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6481 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6541 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6601 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6661 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6721 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6781 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6841 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6901 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6961 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7021 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7081 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7141 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7201 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7381 gccacctgac gtc
//
