LOCUS       sequence-304699-        6483 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6483)
  AUTHORS   Bao J, Lu Z, Joseph JJ, Carabenciov D, Dimond CC, Pang L, Samsel L, 
            McCoy JP Jr, Leclerc J, Nguyen P, Gius D, Sack MN
  TITLE     Characterization of the murine SIRT3 mitochondrial localization 
            sequence and comparison of mitochondrial enrichment and deacetylase 
            activity of long and short SIRT3 isoforms.
  JOURNAL   J Cell Biochem. 2010 May;110(1):238-47.
  PUBMED    20235147
REFERENCE   2  (bases 1 to 6483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biochem."; date: "2010-05"; volume: "110(1)"; pages: "238-47"
COMMENT     Sequence Label: mSIRT3-L
FEATURES             Location/Qualifiers
     source          1..6483
                     /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             1709..1780
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(1840..1857)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1846..2070
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2116..2544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2203..2222)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2413..2434
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2553..2573)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2558..2887
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2738..2873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2800..2819
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2954..3748
                     /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(3008..3027)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3618..3637
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3922..4043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3959..3978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4013..4032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4092..4108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4105..4127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4116..4132
                     /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(4140..4146,4147..4164,4165..4170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4185..4206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4323..4340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4494..5082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4574..4593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5253..6044,6045..6113))
                     /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     5876..5895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6114..6218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6293..6312)
                     /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 actagtccag tgtggtggaa
      721 ttccgtcgtc ctgcagagca tcatggcgct aagcggtcga ctggcattgg ccgcgctcag
      781 actgtggggt ccgggaggtg ggagaaggcc catatccctc tgtgtgggag cctcaggcgg
      841 ctttggaggt ggaggaagca gtgagaagaa gttttctctg caggatgtag ctgagctgct
      901 tcggaccaga gcctgcagta gggtggtggt catggtgggg gccggcatca gcacacccag
      961 tggcatcccg gacttcagat ccccagggag cggcctctac agcaaccttc agcagtatga
     1021 catcccgtac cctgaagcca tctttgaact tggctttttc tttcacaacc ccaagccctt
     1081 tttcatgttg gccaaggagc tgtaccctgg gcactacagg cccaatgtca ctcactactt
     1141 cctgaggctc ctccacgaca aggagctgct tctgcggctc tatacacaga acatcgacgg
     1201 gcttgagaga gcatctggga tccctgcctc aaagctggtt gaagcccacg ggacctttgt
     1261 aacagctaca tgcacggtct gtcgaaggtc cttcccaggg gaagacatat gggctgatgt
     1321 gatggcggac agggtgcccc gctgccctgt ctgtactggc gttgtgaaac ccgacattgt
     1381 gttctttggg gagcagctgc ctgcaaggtt cctactccat atggctgact tcgctttggc
     1441 agatctgcta ctcattcttg ggacctccct ggaggtggag ccttttgcca gcttgtctga
     1501 agcagtacag aaatcagtgc cccgactgct catcaatcga gacttggtgg ggccgttcgt
     1561 tctgagtcct cgaaggaaag atgtggtcca gctaggggat gtagttcatg gtgtggaaag
     1621 gctggtggac ctcctggggt ggacacaaga actgctggat cttatgcagc gggaacgtgg
     1681 caagctggat ggacaggaca gactcgagga ctacaaggat gacgatgaca aggattacaa
     1741 agacgacgat gataaggact ataaggatga tgacgacaaa taatagcaat tcctcgacga
     1801 ctgcatgctc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     1861 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     1921 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1981 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2041 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2101 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2161 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2221 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2281 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2341 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2401 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2461 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2521 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     2581 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2641 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2701 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2761 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2821 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2881 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     2941 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3001 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3061 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3121 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3181 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3241 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3301 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3361 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3421 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3481 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3541 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3601 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3661 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3721 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3781 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3841 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3901 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3961 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4021 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6241 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6301 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6361 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     6421 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     6481 gtt
//
