LOCUS       sequence-198040-        5321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human SIRT6 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-FLAG
FEATURES             Location/Qualifiers
     source          1..5321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        788..1091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1092..1295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1246..1266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1341..1359
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2467..2490
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2543..2562)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2544..2562)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2836..2957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2873..2892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2927..2946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2964..3419)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3101..3122)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3313..3332
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3446..3550
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3552..3909
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3552..3743
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3575..3595)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3760..3895
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3944..4738
                     /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             3944..4738
                     /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(3998..4017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4608..4627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4926..4945)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4970..5017
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      781 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      841 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      901 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      961 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1021 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1081 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     1141 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     1201 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     1261 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
     1321 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac
     1381 cgcggtggcg gccgcatgtc ggtgaattac gcggcggggc tgtcgccgta cgcggacaag
     1441 ggcaagtgcg gcctcccgga gatcttcgac cccccggagg agctggagcg gaaggtgtgg
     1501 gaactggcga ggctggtctg gcagtcttcc agtgtggtgt tccacacggg tgccggcatc
     1561 agcactgcct ctggcatccc cgacttcagg ggtccccacg gagtctggac catggaggag
     1621 cgaggtctgg cccccaagtt cgacaccacc tttgagagcg cgcggcccac gcagacccac
     1681 atggcgctgg tgcagctgga gcgcgtgggc ctcctccgct tcctggtcag ccagaacgtg
     1741 gacgggctcc atgtgcgctc aggcttcccc agggacaaac tggcagagct ccacgggaac
     1801 atgtttgtgg aagaatgtgc caagtgtaag acgcagtacg tccgagacac agtcgtgggc
     1861 accatgggcc tgaaggccac gggccggctc tgcaccgtgg ctaaggcaag ggggctgcga
     1921 gcctgcaggg gagagctgag ggacaccatc ctagactggg aggactccct gcccgaccgg
     1981 gacctggcac tcgccgatga ggccagcagg aacgccgacc tgtccatcac gctgggtaca
     2041 tcgctgcaga tccggcccag cgggaacctg ccgctggcta ccaagcgccg gggaggccgc
     2101 ctggtcatcg tcaacctgca gcccaccaag cacgaccgcc atgctgacct ccgcatccat
     2161 ggctacgttg acgaggtcat gacccggctc atgaagcacc tggggctgga gatccccgcc
     2221 tgggacggcc cccgtgtgct ggagagggcg ctgccacccc tgccccgccc gcccaccccc
     2281 aagctggagc ccaaggagga atctcccacc cggatcaacg gctctatccc cgccggcccc
     2341 aagcaggagc cctgcgccca gcacaacggc tcagagcccg ccagccccaa acgggagcgg
     2401 cccaccagcc ctgcccccca cagacccccc aaaagggtga aggccaaggc ggtccccagc
     2461 ctcgaggatt acaaggatga cgacgataag tagggcccgg taccttaatt aattaaggta
     2521 ccaggtaagt gtacccaatt cgccctatag tgagtcgtat tacaattcac tcgatcgccc
     2581 ttcccaacag ttgcgcagcc tgaatggcga atggagatcc aatttttaag tgtataatgt
     2641 gttaaactac tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc
     2701 aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag
     2761 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2821 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2881 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2941 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3001 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3061 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3121 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3181 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3241 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3301 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3361 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3421 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3481 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3541 aaggaagaat cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3601 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3661 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3721 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3781 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3841 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3901 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3961 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4021 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4081 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4141 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4201 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4261 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4321 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4381 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagaacatc aggggctcgc
     4441 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4501 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4561 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4621 tgatattgct gaagaacttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4681 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4741 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4801 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4861 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4921 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4981 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5041 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5101 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5161 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5221 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5281 atcccttaac gtgagttttc gttccactga gcgtcagacc c
//
