LOCUS       sequence-198051-        8460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human SIRT6 for mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8460)
  AUTHORS   Zhang X, Khan S, Jiang H, Antonyak MA, Chen X, Spiegelman NA, Shrimp
            JH, Cerione RA, Lin H
  TITLE     Identifying the functional contribution of the defatty-acylase 
            activity of SIRT6.
  JOURNAL   Nat Chem Biol. 2016 Aug;12(8):614-20. doi: 10.1038/nchembio.2106. 
            Epub 2016 Jun 20.
  PUBMED    27322069
REFERENCE   2  (bases 1 to 8460)
  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"; doi: 
            "10.1038/nchembio.2106"; journalName: "Nat Chem Biol"; date: 
            "2016-08"; volume: "12"; issue: "8"; pages: "614-20"
COMMENT     Sequence Label: pCDH-SIRT6
FEATURES             Location/Qualifiers
     source          1..8460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..26
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     18..35
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     19..35
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        50..276
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             277..457
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    504..629
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1122..1355
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1540..1584
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1851..1967
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2057..2260
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2211..2231
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2257..2281
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3458..3669
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3682..3950
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3977..4576
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3977..3996)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4473..4493
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4583..5171
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4636..4656)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5054..5065)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5245..5478
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5550..5671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5587..5606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5641..5660
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5711..5846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5879..5895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5879..5895)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5892..5914)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5903..5919
                     /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(5927..5933,5934..5951,5952..5957))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5972..5993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6110..6127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6281..6869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7040..7831,7832..7900))
                     /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     7663..7682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7901..8005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8073..8091
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8129..8151)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8251..8270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctgacgcgt gtagtcttat
       61 gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga
      121 gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta
      181 ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga
      241 gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc
      301 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      361 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      421 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      481 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      541 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      601 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      661 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      721 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      781 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      841 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      901 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      961 agcaagcggc cactgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1021 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1081 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1141 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1201 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1261 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1321 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1381 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1441 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1501 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1561 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1621 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1681 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1741 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1801 agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga
     1861 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     1921 atacaaacta aagaattaca aaaacaaatt acaaaattca aaattttatc gatactagta
     1981 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2041 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2101 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2161 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2221 cggtaggcgt gtacggtggg aggtttatat aagcagagct cgtttagtga accgtcagat
     2281 cgcctggaga cgccatccac gctgttttga cctccataga agattctaga gctagcgaat
     2341 tcatgtcggt gaattacgcg gcggggctgt cgccgtacgc ggacaagggc aagtgcggcc
     2401 tcccggagat cttcgacccc ccggaggagc tggagcggaa ggtgtgggaa ctggcgaggc
     2461 tggtctggca gtcttccagt gtggtgttcc acacgggtgc cggcatcagc actgcctctg
     2521 gcatccccga cttcaggggt ccccacggag tctggaccat ggaggagcga ggtctggccc
     2581 ccaagttcga caccaccttt gagagcgcgc ggcccacgca gacccacatg gcgctggtgc
     2641 agctggagcg cgtgggcctc ctccgcttcc tggtcagcca gaacgtggac gggctccatg
     2701 tgcgctcagg cttccccagg gacaaactgg cagagctcca cgggaacatg tttgtggaag
     2761 aatgtgccaa gtgtaagacg cagtacgtcc gagacacagt cgtgggcacc atgggcctga
     2821 aggccacggg ccggctctgc accgtggcta aggcaagggg gctgcgagcc tgcaggggag
     2881 agctgaggga caccatccta gactgggagg actccctgcc cgaccgggac ctggcactcg
     2941 ccgatgaggc cagcaggaac gccgacctgt ccatcacgct gggtacatcg ctgcagatcc
     3001 ggcccagcgg gaacctgccg ctggctacca agcgccgggg aggccgcctg gtcatcgtca
     3061 acctgcagcc caccaagcac gaccgccatg ctgacctccg catccatggc tacgttgacg
     3121 aggtcatgac ccggctcatg aagcacctgg ggctggagat ccccgcctgg gacggccccc
     3181 gtgtgctgga gagggcgctg ccacccctgc cccgcccgcc cacccccaag ctggagccca
     3241 aggaggaatc tcccacccgg atcaacggct ctatccccgc cggccccaag caggagccct
     3301 gcgcccagca caacggctca gagcccgcca gccccaaacg ggagcggccc accagccctg
     3361 ccccccacag accccccaaa agggtgaagg ccaaggcggt ccccagctga ggatccgcgg
     3421 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     3481 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc
     3541 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     3601 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     3661 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc
     3721 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc
     3781 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg
     3841 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt
     3901 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga
     3961 ccggcgccta cgctagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4021 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4081 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4141 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4201 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4261 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4321 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4381 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4441 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4501 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4561 caagcccggt gcctgagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4621 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4681 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4741 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4801 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4861 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4921 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4981 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5041 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5101 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5161 cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5221 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agataagatc
     5281 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5341 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5401 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5461 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     5521 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     5581 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5641 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     5701 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5761 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5821 gtagtgagga ggcttttttg gaggcctaga cttttgcaga gaccaaattc gtaatcatgt
     5881 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5941 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6001 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6061 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8041 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8101 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     8161 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     8221 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     8281 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     8341 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     8401 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
//
