LOCUS       sequence-198056-        8460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human SIRT6 H133Y 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-H133Y
FEATURES             Location/Qualifiers
     source          1..8460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15..31
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        46..272
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             273..453
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    500..625
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1118..1351
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1536..1580
                     /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    1847..1963
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2053..2256
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2207..2227
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2253..2277
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3454..3665
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3678..3946
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3973..4572
                     /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(3973..3992)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4469..4489
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4579..5167
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4632..4652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5050..5061)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5241..5474
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5546..5667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5583..5602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5637..5656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5707..5842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5875..5891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5875..5891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5888..5910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5899..5915
                     /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(5923..5929,5930..5947,5948..5953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5968..5989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6106..6123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6277..6865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7036..7827,7828..7896))
                     /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     7659..7678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7897..8001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8069..8087
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8125..8147)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8247..8266
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(8460,1..22)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg acgcgtgtag tcttatgcaa
       61 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
      121 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
      181 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
      241 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
      301 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      361 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      421 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      481 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      541 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      601 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      661 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      721 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      781 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
      841 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
      901 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
      961 agcggccact gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     1021 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     1081 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     1141 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     1201 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     1261 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     1321 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     1381 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     1441 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     1501 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     1561 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     1621 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     1681 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     1741 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     1801 acagatccat tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag
     1861 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     1921 aaactaaaga attacaaaaa caaattacaa aattcaaaat tttatcgata ctagtattat
     1981 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2041 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2101 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2161 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2221 aggcgtgtac ggtgggaggt ttatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2281 tggagacgcc atccacgctg ttttgacctc catagaagat tctagagcta gcgaattcat
     2341 gtcggtgaat tacgcggcgg ggctgtcgcc gtacgcggac aagggcaagt gcggcctccc
     2401 ggagatcttc gaccccccgg aggagctgga gcggaaggtg tgggaactgg cgaggctggt
     2461 ctggcagtct tccagtgtgg tgttccacac gggtgccggc atcagcactg cctctggcat
     2521 ccccgacttc aggggtcccc acggagtctg gaccatggag gagcgaggtc tggcccccaa
     2581 gttcgacacc acctttgaga gcgcgcggcc cacgcagacc cacatggcgc tggtgcagct
     2641 ggagcgcgtg ggcctcctcc gcttcctggt cagccagaac gtggacgggc tccatgtgcg
     2701 ctcaggcttc cccagggaca aactggcaga gctctacggg aacatgtttg tggaagaatg
     2761 tgccaagtgt aagacgcagt acgtccgaga cacagtcgtg ggcaccatgg gcctgaaggc
     2821 cacgggccgg ctctgcaccg tggctaaggc aagggggctg cgagcctgca ggggagagct
     2881 gagggacacc atcctagact gggaggactc cctgcccgac cgggacctgg cactcgccga
     2941 tgaggccagc aggaacgccg acctgtccat cacgctgggt acatcgctgc agatccggcc
     3001 cagcgggaac ctgccgctgg ctaccaagcg ccggggaggc cgcctggtca tcgtcaacct
     3061 gcagcccacc aagcacgacc gccatgctga cctccgcatc catggctacg ttgacgaggt
     3121 catgacccgg ctcatgaagc acctggggct ggagatcccc gcctgggacg gcccccgtgt
     3181 gctggagagg gcgctgccac ccctgccccg cccgcccacc cccaagctgg agcccaagga
     3241 ggaatctccc acccggatca acggctctat ccccgccggc cccaagcagg agccctgcgc
     3301 ccagcacaac ggctcagagc ccgccagccc caaacgggag cggcccacca gccctgcccc
     3361 ccacagaccc cccaaaaggg tgaaggccaa ggcggtcccc agctgaggat ccgcggccgc
     3421 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     3481 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
     3541 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     3601 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     3661 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     3721 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     3781 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     3841 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     3901 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     3961 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4021 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4081 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4141 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4201 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4261 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4321 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4381 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4441 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4501 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4561 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4621 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4681 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4741 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4801 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4861 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4921 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     4981 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5041 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5101 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5161 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5221 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct
     5281 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5341 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5401 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     5461 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     5521 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5641 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     5701 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5761 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5821 tgaggaggct tttttggagg cctagacttt tgcagagacc aaattcgtaa tcatgtcata
     5881 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5941 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6001 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6061 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8041 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8101 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     8161 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     8221 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     8281 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     8341 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8401 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
//
