LOCUS       sequence-198054-        8460 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Human SIRT6 G60A 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 Apr 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-G60A
FEATURES             Location/Qualifiers
     source          1..8460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..21
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5..21
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        36..262
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             263..443
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    490..615
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1108..1341
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1526..1570
                     /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    1837..1953
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2043..2246
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2197..2217
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2243..2267
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3444..3655
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3668..3936
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3963..4562
                     /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(3963..3982)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4459..4479
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4569..5157
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4622..4642)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5040..5051)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5231..5464
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5536..5657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5573..5592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5627..5646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5697..5832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5865..5881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5865..5881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5878..5900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5889..5905
                     /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(5913..5919,5920..5937,5938..5943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5958..5979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6096..6113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6267..6855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7026..7817,7818..7886))
                     /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     7649..7668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7887..7991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8059..8077
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8115..8137)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8237..8256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(8450..8460,1..12)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgttgtaaaa cgacggccag tgccaagctg acgcgtgtag tcttatgcaa tactcttgta
       61 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
      121 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
      181 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
      241 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
      301 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      361 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
      421 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
      481 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
      541 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
      601 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
      661 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
      721 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
      781 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
      841 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
      901 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccact
      961 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1021 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1081 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1141 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1201 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1261 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1321 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     1381 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     1441 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     1501 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     1561 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     1621 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     1681 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     1741 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     1801 tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag gggggattgg
     1861 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     1921 attacaaaaa caaattacaa aattcaaaat tttatcgata ctagtattat gcccagtaca
     1981 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2041 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     2101 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     2161 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     2221 ggtgggaggt ttatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     2281 atccacgctg ttttgacctc catagaagat tctagagcta gcgaattcat gtcggtgaat
     2341 tacgcggcgg ggctgtcgcc gtacgcggac aagggcaagt gcggcctccc ggagatcttc
     2401 gaccccccgg aggagctgga gcggaaggtg tgggaactgg cgaggctggt ctggcagtct
     2461 tccagtgtgg tgttccacac gggtgccggc atcagcactg cctctgccat ccccgacttc
     2521 aggggtcccc acggagtctg gaccatggag gagcgaggtc tggcccccaa gttcgacacc
     2581 acctttgaga gcgcgcggcc cacgcagacc cacatggcgc tggtgcagct ggagcgcgtg
     2641 ggcctcctcc gcttcctggt cagccagaac gtggacgggc tccatgtgcg ctcaggcttc
     2701 cccagggaca aactggcaga gctccacggg aacatgtttg tggaagaatg tgccaagtgt
     2761 aagacgcagt acgtccgaga cacagtcgtg ggcaccatgg gcctgaaggc cacgggccgg
     2821 ctctgcaccg tggctaaggc aagggggctg cgagcctgca ggggagagct gagggacacc
     2881 atcctagact gggaggactc cctgcccgac cgggacctgg cactcgccga tgaggccagc
     2941 aggaacgccg acctgtccat cacgctgggt acatcgctgc agatccggcc cagcgggaac
     3001 ctgccgctgg ctaccaagcg ccggggaggc cgcctggtca tcgtcaacct gcagcccacc
     3061 aagcacgacc gccatgctga cctccgcatc catggctacg ttgacgaggt catgacccgg
     3121 ctcatgaagc acctggggct ggagatcccc gcctgggacg gcccccgtgt gctggagagg
     3181 gcgctgccac ccctgccccg cccgcccacc cccaagctgg agcccaagga ggaatctccc
     3241 acccggatca acggctctat ccccgccggc cccaagcagg agccctgcgc ccagcacaac
     3301 ggctcagagc ccgccagccc caaacgggag cggcccacca gccctgcccc ccacagaccc
     3361 cccaaaaggg tgaaggccaa ggcggtcccc agctgaggat ccgcggccgc gaaggatctg
     3421 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3481 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     3541 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     3601 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     3661 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     3721 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     3781 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     3841 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     3901 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     3961 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     4021 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     4081 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     4141 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     4201 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4261 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4321 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4381 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4441 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4501 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4561 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     4621 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     4681 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     4741 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     4801 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     4861 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4921 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     4981 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     5041 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     5101 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     5161 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5221 aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct ttttgcttgt
     5281 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5341 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5401 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5461 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     5521 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5581 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5641 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     5701 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5761 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5821 tttttggagg cctagacttt tgcagagacc aaattcgtaa tcatgtcata gctgtttcct
     5881 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5941 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6001 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7981 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8041 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     8101 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     8161 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     8221 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     8281 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     8341 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8401 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
//
