LOCUS       sequence-198047-        5321 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Human SIRT6 G60A 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 Apr 14, 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-G60A-FLAG
FEATURES             Location/Qualifiers
     source          1..5321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        782..1085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1086..1289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1240..1260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1335..1353
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2461..2484
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2537..2556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2538..2556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2830..2951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2867..2886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2921..2940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2958..3413)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3095..3116)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3307..3326
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3440..3544
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3546..3903
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3546..3737
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3569..3589)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3754..3889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3938..4732
                     /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             3938..4732
                     /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(3992..4011)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4602..4621
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4920..4939)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4964..5011
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ccctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
      721 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      781 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      841 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      901 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      961 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     1021 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1081 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1141 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1201 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1261 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgat
     1321 tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt
     1381 ggcggccgca tgtcggtgaa ttacgcggcg gggctgtcgc cgtacgcgga caagggcaag
     1441 tgcggcctcc cggagatctt cgaccccccg gaggagctgg agcggaaggt gtgggaactg
     1501 gcgaggctgg tctggcagtc ttccagtgtg gtgttccaca cgggtgccgg catcagcact
     1561 gcctctgcca tccccgactt caggggtccc cacggagtct ggaccatgga ggagcgaggt
     1621 ctggccccca agttcgacac cacctttgag agcgcgcggc ccacgcagac ccacatggcg
     1681 ctggtgcagc tggagcgcgt gggcctcctc cgcttcctgg tcagccagaa cgtggacggg
     1741 ctccatgtgc gctcaggctt ccccagggac aaactggcag agctccacgg gaacatgttt
     1801 gtggaagaat gtgccaagtg taagacgcag tacgtccgag acacagtcgt gggcaccatg
     1861 ggcctgaagg ccacgggccg gctctgcacc gtggctaagg caagggggct gcgagcctgc
     1921 aggggagagc tgagggacac catcctagac tgggaggact ccctgcccga ccgggacctg
     1981 gcactcgccg atgaggccag caggaacgcc gacctgtcca tcacgctggg tacatcgctg
     2041 cagatccggc ccagcgggaa cctgccgctg gctaccaagc gccggggagg ccgcctggtc
     2101 atcgtcaacc tgcagcccac caagcacgac cgccatgctg acctccgcat ccatggctac
     2161 gttgacgagg tcatgacccg gctcatgaag cacctggggc tggagatccc cgcctgggac
     2221 ggcccccgtg tgctggagag ggcgctgcca cccctgcccc gcccgcccac ccccaagctg
     2281 gagcccaagg aggaatctcc cacccggatc aacggctcta tccccgccgg ccccaagcag
     2341 gagccctgcg cccagcacaa cggctcagag cccgccagcc ccaaacggga gcggcccacc
     2401 agccctgccc cccacagacc ccccaaaagg gtgaaggcca aggcggtccc cagcctcgag
     2461 gattacaagg atgacgacga taagtagggc ccggtacctt aattaattaa ggtaccaggt
     2521 aagtgtaccc aattcgccct atagtgagtc gtattacaat tcactcgatc gcccttccca
     2581 acagttgcgc agcctgaatg gcgaatggag atccaatttt taagtgtata atgtgttaaa
     2641 ctactgattc taattgtttg tgtattttag attcacagtc ccaaggctca tttcaggccc
     2701 ctcagtcctc acagtctgtt catgatcata atcagccata ccacatttgt agaggtttta
     2761 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2821 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2881 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2941 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3001 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3061 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3121 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3181 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3241 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3301 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3361 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3541 gaatcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3601 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3781 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3841 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3901 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3961 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4021 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4081 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4141 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4201 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4261 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4321 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4381 ggaagccggt cttgtcgatc aggatgatct ggacgaagaa catcaggggc tcgcgccagc
     4441 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4501 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4561 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4621 tgctgaagaa cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4681 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4741 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4801 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4861 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4921 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4981 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5041 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5101 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5161 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag a
//
