LOCUS       sequence-137158-        6486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human SIRT6 with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  AUTHORS   North BJ, Marshall BL, Borra MT, Denu JM, Verdin E
  TITLE     The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin 
            deacetylase.
  JOURNAL   Mol Cell. 2003 Feb . 11(2):437-44.
  PUBMED    12620231
REFERENCE   2  (bases 1 to 6486)
  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"; journalName: "Mol Cell. 
            2003 Feb . 11(2):437-44."
COMMENT     Sequence Label: SIRT6 Flag
FEATURES             Location/Qualifiers
     source          1..6486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1785..1808
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1843..1860)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1849..2073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2119..2547
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2206..2225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2416..2437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2556..2576)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2561..2890
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2741..2876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2803..2822
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2957..3751
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3011..3030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3621..3640
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3925..4046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3962..3981)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4016..4035
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4095..4111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4095..4111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4108..4130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4119..4135
                     /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(4143..4149,4150..4167,4168..4173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4188..4209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4326..4343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4497..5085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4577..4596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5256..6047,6048..6116))
                     /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     5879..5898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6117..6221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6296..6315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gaattccatg tcggtgaatt
      721 acgcggcggg gctgtcgccg tacgcggaca agggcaagtg cggcctcccg gagatcttcg
      781 accccccgga ggagctggag cggaaggtgt gggaactggc gaggctggtc tggcagtctt
      841 ccaatgtggt gttccacacg ggtgccggca tcagcactgc ctctggcatc cccgacttca
      901 ggggtcccca cggagtctgg accatggagg agcgaggtct ggcccccaag ttcgacacca
      961 cctttgagag cgcgcggccc acgcagaccc acatggcgct ggtgcagctg gagcgcgtgg
     1021 gcctcctccg cttcctggtc agccagaacg tggacgggct ccatgtgcgc tcaggcttcc
     1081 ccagggacaa actggcagag ctccacggga acatgtttgt ggaagaatgt gccaagtgta
     1141 agacgcagta cgtccgagac acagtcgtgg gcaccatggg cctgaaggcc acgggccggc
     1201 tctgcaccgt ggctaaggca agggggctgc gagcctgcag gggagagctg agggacacca
     1261 tcctagactg ggaggactcc ctgcccgacc gggacctggc actcgccgat gaggccagca
     1321 ggaacgccga cctgtccatc acgctgggta catcgctgca gatccggccc agcgggaacc
     1381 tgccgctggc taccaagcgc cggggaggcc gcctggtcat cgtcaacctg cagcccacca
     1441 agcacgaccg ccatgctgac ctccgcatcc atggctacgt tgacgaggtc atgacccggc
     1501 tcatgaagca cctggggctg gagatccccg cctgggacgg cccccgtgtg ctggagaggg
     1561 cgctgccacc cctgccccgc ccgcccaccc ccaagctgga gcccaaggag gaatctccca
     1621 cccggatcaa cggctctatc cccgccggcc ccaagcagga gccctgcgcc cagcacaacg
     1681 gctcagagcc cgccagcccc aaacgggagc ggcccaccag ccctgccccc cacagacccc
     1741 ccaaaagggt gaaggccaag gcggtcccca gcgaattcgg aggagactac aaggacgacg
     1801 atgacaagta atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     1861 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1921 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1981 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2041 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2101 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2161 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2221 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2281 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2341 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2401 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2461 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2521 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     2581 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2641 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2701 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2761 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2821 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2881 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     2941 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3001 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3061 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3121 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3181 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3241 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3301 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3361 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3421 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3481 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3541 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3601 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3661 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3721 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3781 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3841 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3901 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3961 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4021 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4201 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4261 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4921 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5161 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5221 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5281 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5341 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5401 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5461 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5521 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5581 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5641 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5701 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5761 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5821 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5881 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5941 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6001 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6061 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6241 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6301 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6361 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     6421 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     6481 cgcgtt
//
