LOCUS       sequence-272724-        6621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6621)
  AUTHORS   Hirschey MD, Shimazu T, Goetzman E, Jing E, Schwer B, Lombard DB, 
            Grueter CA, Harris C, Biddinger S, Ilkayeva OR, Stevens RD, Li Y, 
            Saha AK, Ruderman NB, Bain JR, Newgard CB, Farese RV, Alt FW, Kahn 
            CR, Verdin E
  TITLE     SIRT3 regulates mitochondrial fatty-acid oxidation by reversible 
            enzyme deacetylation.
  JOURNAL   Nature. 2010 Mar 4. 464(7285):121-5.
  PUBMED    20203611
REFERENCE   2  (bases 1 to 6621)
  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: "Nature. 
            2010 Mar 4. 464(7285):121-5."
COMMENT     Sequence Label: pcDNA3.1hSIRT3_H248Y_HA
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1917..1943
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1978..1995)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1984..2208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2254..2682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2341..2360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2551..2572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2691..2711)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2696..3025
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2876..3011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2938..2957
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3092..3886
                     /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(3146..3165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3756..3775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4060..4181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4097..4116)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4151..4170
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4230..4246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4230..4246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4243..4265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4254..4270
                     /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(4278..4284,4285..4302,4303..4308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4323..4344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4461..4478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4632..5220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4712..4731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5391..6182,6183..6251))
                     /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     6014..6033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6252..6356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6431..6450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gaattccatg gcgttctggg
      721 gttggcgcgc cgcggcagcc ctccggctgt ggggccgggt agttgaacgg gtcgaggccg
      781 ggggaggcgt ggggccgttt catgcctgcg gctgtcggct ggtgcttggc ggcagggacg
      841 atgtgagtgc ggggctgaga ggcagccatg gggcccgcgg tgagcccttg gacccggcgc
      901 gccccttgca gaggcctccc agacccgagg tgcccagggc attccggagg cagccgaggg
      961 cagcagctcc cagtttcttc ttttcgagta ttaaaggtgg aagaaggtcc atatcttttt
     1021 ctgtgggtgc ttcaagtgtt gttggaagtg gaggcagcag tgacaagggg aagctttccc
     1081 tgcaggatgt agctgagctg attcgggcca gagcctgcca gagggtggtg gtcatggtgg
     1141 gggccggcat cagcacaccc agtggcattc cagacttcag atcgccgggg agtggcctgt
     1201 acagcaacct ccagcagtac gatctcccgt accccgaggc catttttgaa ctcccattct
     1261 tctttcacaa ccccaagccc tttttcactt tggccaagga gctgtaccct ggaaactaca
     1321 agcccaacgt cactcactac tttctccggc tgcttcatga caaggggctg cttctgcggc
     1381 tctacacgca gaacatcgat gggcttgaga gagtgtcggg catccctgcc tcaaagctgg
     1441 ttgaagctta tggaaccttt gcctctgcca cctgcacagt ctgccaaaga cccttcccag
     1501 gggaggacat tcgggctgac gtgatggcag acagggttcc ccgctgcccg gtctgcaccg
     1561 gcgttgtgaa gcccgacatt gtgttctttg gggagccgct gccccagagg ttcttgctgc
     1621 atgtggttga tttccccatg gcagatctgc tgctcatcct tgggacctcc ctggaggtgg
     1681 agccttttgc cagcttgacc gaggccgtgc ggagctcagt tccccgactg ctcatcaacc
     1741 gggacttggt ggggcccttg gcttggcatc ctcgcagcag ggacgtggcc cagctggggg
     1801 acgtggttca cggcgtggaa agcctagtgg agcttctggg ctggacagaa gagatgcggg
     1861 accttgtgca gcgggaaact gggaagcttg atggaccaga caaagaattc ggaggatacc
     1921 catacgacgt cccagactac gcttaatcta gagggcccgt ttaaacccgc tgatcagcct
     1981 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2041 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2101 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2161 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2221 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2281 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2341 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2401 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2461 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2521 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2581 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2641 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2701 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2761 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2821 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2881 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2941 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3001 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3061 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3121 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3181 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3241 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3301 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3361 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3421 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3481 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3541 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3601 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3661 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3721 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3781 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3841 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3901 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3961 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4021 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4081 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4141 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4201 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4261 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4321 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4381 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4441 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4501 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4561 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4621 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4681 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4741 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4801 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4861 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4921 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4981 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5041 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5101 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5161 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5221 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5281 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5341 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5401 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5461 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5521 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5581 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5641 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5701 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5761 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5821 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5881 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5941 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6001 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6061 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6121 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6181 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6241 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6301 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6361 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6421 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6481 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6541 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     6601 tacgggccag atatacgcgt t
//
