LOCUS       sequence-238527-        9058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human histone deacetylase 6 with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9058)
  AUTHORS   Fischle W, Emiliani S, Hendzel MJ, Nagase T, Nomura N, Voelter W, 
            Verdin E
  TITLE     A new family of human histone deacetylases related to Saccharomyces 
            cerevisiae HDA1p.
  JOURNAL   J Biol Chem. 1999 Apr 23. 274(17):11713-20.
  PUBMED    10206986
REFERENCE   2  (bases 1 to 9058)
  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: "J Biol 
            Chem. 1999 Apr 23. 274(17):11713-20."
COMMENT     Sequence Label: HDAC6 Flag
FEATURES             Location/Qualifiers
     source          1..9058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4591..4614
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4649..4666)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4655..4879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4925..5353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5012..5031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5222..5243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5362..5382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5367..5696
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5547..5682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5609..5628
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5763..6557
                     /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(5817..5836)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6427..6446
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6731..6852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6768..6787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6822..6841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6901..6917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6901..6917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6914..6936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6925..6941
                     /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(6949..6955,6956..6973,6974..6979))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6994..7015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7132..7149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7303..7891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7383..7402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8062..8853,8854..8922))
                     /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     8685..8704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8923..9027)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcgg ccgctcaact atgacctcaa ccggccagga ttccaccaca
      961 accaggcagc gaagaagtag gcagaacccc cagtcgcccc ctcaggactc cagtgtcact
     1021 tcgaagcgaa atattaaaaa gggagccgtt ccccgctcta tccccaatct agcggaggta
     1081 aagaagaaag gcaaaatgaa gaagctcggc caagcaatgg aagaagacct aatcgtggga
     1141 ctgcaaggga tggatctgaa ccttgaggct gaagcactgg ctggcactgg cttggtgttg
     1201 gatgagcagt taaatgaatt ccattgcctc tgggatgaca gcttcccgga aggccctgag
     1261 cggctccatg ccatcaagga gcaactgatc caggagggcc tcctagatcg ctgcgtgtcc
     1321 tttcaggccc ggtttgctga aaaggaagag ctgatgttgg ttcacagcct agaatatatt
     1381 gatctgatgg aaacaaccca gtacatgaat gagggagaac tccgtgtcct agcagacacc
     1441 tacgactcag tttatctgca tccgaactca tactcctgtg cctgcctggc ctcaggctct
     1501 gtcctcaggc tggtggatgc ggtcctgggg gctgagatcc ggaatggcat ggccatcatt
     1561 aggcctcctg gacatcacgc ccagcacagt cttatggatg gctattgcat gttcaaccac
     1621 gtggctgtgg cagcccgcta tgctcaacag aaacaccgca tccggagggt ccttatcgta
     1681 gattgggatg tgcaccacgg tcaaggaaca cagttcacct tcgaccagga ccccagtgtc
     1741 ctctatttct ccatccaccg ctacgagcag ggtaggttct ggccccacct gaaggcctct
     1801 aactggtcca ccacaggttt cggccaaggc caaggatata ccatcaatgt gccttggaac
     1861 caggtgggga tgcgggatgc tgactacatt gctgctttcc tgcacgtcct gctgccagtc
     1921 gccctcgagt tccagcctca gctggtcctg gtggctgctg gatttgatgc cctgcaaggg
     1981 gaccccaagg gtgagatggc cgccactccg gcagggttcg cccagctaac ccacctgctc
     2041 atgggtctgg caggaggcaa gctgatcctg tctctggagg gtggctacaa cctccgcgcc
     2101 ctggctgaag gcgtcagtgc ttcgctccac acccttctgg gagacccttg ccccatgctg
     2161 gagtcacctg gtgccccctg ccggagtgcc caggcttcag tttcctgtgc tctggaagcc
     2221 cttgagccct tctgggaggt tcttgtgaga tcaactgaga ccgtggagag ggacaacatg
     2281 gaggaggaca atgtagagga gagcgaggag gaaggaccct gggagccccc tgtgctccca
     2341 atcctgacat ggccagtgct acagtctcgc acagggctgg tctatgacca aaatatgatg
     2401 aatcactgca acttgtggga cagccaccac cctgaggtac cccagcgcat cttgcggatc
     2461 atgtgccgtc tggaggagct gggccttgcc gggcgctgcc tcaccctgac accgcgccct
     2521 gccacagagg ctgagctgct cacctgtcac agtgctgagt acgtgggtca tctccgggcc
     2581 acagagaaaa tgaaaacccg ggagctgcac cgtgagagtt ccaactttga ctccatctat
     2641 atctgcccca gtaccttcgc ctgtgcacag cttgccactg gcgctgcctg ccgcctggtg
     2701 gaggctgtgc tctcaggaga ggttctgaat ggtgctgctg tggtgcgtcc cccaggacac
     2761 cacgcagagc aggatgcagc ttgcggtttt tgctttttca actctgtggc tgtggctgct
     2821 cgccatgccc agactatcag tgggcatgcc ctacggatcc tgattgtgga ttgggatgtc
     2881 caccacggta atggaactca gcacatgttt gaggatgacc ccagtgtgct atatgtgtcc
     2941 ctgcaccgct atgatcatgg caccttcttc cccatggggg atgagggtgc cagcagccag
     3001 atcggccggg ctgcgggcac aggcttcacc gtcaacgtgg catggaacgg gccccgcatg
     3061 ggtgatgctg actacctagc tgcctggcat cgcctggtgc ttcccattgc ctacgagttt
     3121 aacccagaac tggtgctggt ctcagctggc tttgatgctg cacgggggga tccgctgggg
     3181 ggctgccagg tgtcacctga gggttatgcc cacctcaccc acctgctgat gggccttgcc
     3241 agtggccgca ttatccttat cctagagggt ggctataacc tgacatccat ctcagagtcc
     3301 atggctgcct gcactcgctc cctccttgga gacccaccac ccctgctgac cctgccacgg
     3361 cccccactat caggggccct ggcctcaatc actgagacca tccaagtcca tcgcagatac
     3421 tggcgcagct tacgggtcat gaaggtagaa gacagagaag gaccctccag ttctaagttg
     3481 gtcaccaaga aggcacccca accagccaaa cctaggttag ctgagcggat gaccacacga
     3541 gaaaagaagg ttctggaagc aggcatgggg aaagtcacct cggcatcatt tggggaagag
     3601 tccactccag gccagactaa ctcagagaca gctgtggtgg ccctcactca ggaccagccc
     3661 tcagaggcag ccacaggggg agccactctg gcccagacca tttctgaggc agccattggg
     3721 ggagccatgc tgggccagac cacctcagag gaggctgtcg ggggagccac tccggaccag
     3781 accacctcag aggagactgt gggaggagcc attctggacc agaccacctc agaggatgct
     3841 gttgggggag ccacgctggg ccagactacc tcagaggagg ctgtaggagg agctacactg
     3901 gcccagacca cctcggaggc agccatggag ggagccacac tggaccagac tacgtcagag
     3961 gaggctccag ggggcaccga gctgatccaa actcctctag cctcgagcac agaccaccag
     4021 acccccccaa cctcacctgt gcagggaact acaccccaga tatctcccag tacactgatt
     4081 gggagtctca ggaccttgga gctaggcagc gaatctcagg gggcctcaga atctcaggcc
     4141 ccaggagagg agaacctact aggagaggca gctggaggtc aggacatggc tgattcgatg
     4201 ctgatgcagg gatctagggg cctcactgat caggccatat tttatgctgt gacaccactg
     4261 ccctggtgtc cccatttggt ggcagtatgc cccatacctg cagcaggcct agacgtgacc
     4321 caaccttgtg gggactgtgg aacaatccaa gagaattggg tgtgtctctc ttgctatcag
     4381 gtctactgtg gtcgttacat caatggccac atgctccaac accatggaaa ttctggacac
     4441 ccgctggtcc tcagctacat cgacctgtca gcctggtgtt actactgtca ggcctatgtc
     4501 caccaccagg ctctcctaga tgtgaagaac atcgcccacc agaacaagtt tggggaggat
     4561 atgccccacc cacactgcgg ccgcggagga gactacaagg acgacgatga caagtaatct
     4621 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4681 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4741 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4801 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4861 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4921 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4981 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5041 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5101 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5161 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5221 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5281 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5341 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5401 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     5461 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5521 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5581 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5641 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5701 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5761 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5821 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5881 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5941 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6001 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6061 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6121 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6181 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6241 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6301 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6361 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6421 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6481 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6541 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6601 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6661 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6721 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6781 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6841 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6901 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6961 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7021 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7081 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7141 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7201 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7261 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7321 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7381 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7441 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7501 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7561 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7621 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7681 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7741 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7801 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7861 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7921 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7981 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8041 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8101 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8161 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8221 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8281 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8341 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8401 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8461 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8521 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8581 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8641 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8701 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8761 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8821 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8881 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8941 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9001 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
