LOCUS       sequence-198049-        5321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human SIRT6 H133Y with FLAG tag for mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  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 5321)
  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: pCMV4a-SIRT6-H133Y-FLAG
FEATURES             Location/Qualifiers
     source          1..5321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        778..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1331..1349
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2457..2480
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2533..2552)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2534..2552)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2826..2947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2863..2882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2917..2936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2954..3409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3091..3112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3303..3322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3436..3540
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3542..3899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3542..3733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3565..3585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3750..3885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3934..4728
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3934..4728
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3988..4007)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4598..4617
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4916..4935)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4960..5007
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgattacg
     1321 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
     1381 gccgcatgtc ggtgaattac gcggcggggc tgtcgccgta cgcggacaag ggcaagtgcg
     1441 gcctcccgga gatcttcgac cccccggagg agctggagcg gaaggtgtgg gaactggcga
     1501 ggctggtctg gcagtcttcc agtgtggtgt tccacacggg tgccggcatc agcactgcct
     1561 ctggcatccc cgacttcagg ggtccccacg gagtctggac catggaggag cgaggtctgg
     1621 cccccaagtt cgacaccacc tttgagagcg cgcggcccac gcagacccac atggcgctgg
     1681 tgcagctgga gcgcgtgggc ctcctccgct tcctggtcag ccagaacgtg gacgggctcc
     1741 atgtgcgctc aggcttcccc agggacaaac tggcagagct ctacgggaac atgtttgtgg
     1801 aagaatgtgc caagtgtaag acgcagtacg tccgagacac agtcgtgggc accatgggcc
     1861 tgaaggccac gggccggctc tgcaccgtgg ctaaggcaag ggggctgcga gcctgcaggg
     1921 gagagctgag ggacaccatc ctagactggg aggactccct gcccgaccgg gacctggcac
     1981 tcgccgatga ggccagcagg aacgccgacc tgtccatcac gctgggtaca tcgctgcaga
     2041 tccggcccag cgggaacctg ccgctggcta ccaagcgccg gggaggccgc ctggtcatcg
     2101 tcaacctgca gcccaccaag cacgaccgcc atgctgacct ccgcatccat ggctacgttg
     2161 acgaggtcat gacccggctc atgaagcacc tggggctgga gatccccgcc tgggacggcc
     2221 cccgtgtgct ggagagggcg ctgccacccc tgccccgccc gcccaccccc aagctggagc
     2281 ccaaggagga atctcccacc cggatcaacg gctctatccc cgccggcccc aagcaggagc
     2341 cctgcgccca gcacaacggc tcagagcccg ccagccccaa acgggagcgg cccaccagcc
     2401 ctgcccccca cagacccccc aaaagggtga aggccaaggc ggtccccagc ctcgaggatt
     2461 acaaggatga cgacgataag tagggcccgg taccttaatt aattaaggta ccaggtaagt
     2521 gtacccaatt cgccctatag tgagtcgtat tacaattcac tcgatcgccc ttcccaacag
     2581 ttgcgcagcc tgaatggcga atggagatcc aatttttaag tgtataatgt gttaaactac
     2641 tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca
     2701 gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag gttttacttg
     2761 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2821 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2881 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2941 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3001 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3061 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3121 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3181 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3241 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3301 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3361 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3421 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3481 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat
     3541 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3661 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3721 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3901 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3961 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4021 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4081 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4141 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4201 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4261 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4321 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4381 gccggtcttg tcgatcagga tgatctggac gaagaacatc aggggctcgc gccagccgaa
     4441 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4501 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4561 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4621 gaagaacttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4681 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4741 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4801 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4861 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4921 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4981 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5041 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5101 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5161 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5221 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5281 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa g
//
