LOCUS       Exported                8774 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of human histone deacetylase 5 with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    HDAC5 Flag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8774)
  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 8774)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8774
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4309..4332
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4367..4384)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4373..4597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4643..5071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4730..4749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4940..4961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5080..5100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5085..5414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5265..5400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5327..5346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5481..6275
                     /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(5535..5554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6145..6164
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6449..6570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6486..6505)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6540..6559
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6619..6635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6619..6635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6632..6654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6643..6659
                     /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(6667..6697)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6712..6733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6850..6867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7021..7609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7101..7120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7780..8640)
                     /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     8403..8422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8641..8745)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgc ggccgccacc atgaactctc ccaacgagtc ggatgggatg
      961 tcaggtcggg aaccatcctt ggaaatcctg ccgcggactt ctctgcacag catccctgtg
     1021 acagtggagg tgaagccggt gctgccaaga gccatgccca gttccatggg gggtgggggt
     1081 ggaggcagcc ccagccctgt ggagctacgg ggggctctgg tgggctctgt ggaccccaca
     1141 ctgcgggagc agcaactgca gcaggagctc ctggcgctca agcagcagca gcagctgcag
     1201 aagcagctcc tgttcgctga gttccagaaa cagcatgacc acctgacaag gcagcatgag
     1261 gtccagctgc agaagcacct caagcagcag caggagatgc tggcagccaa gcagcagcag
     1321 gagatgctgg cagccaagcg gcagcaggag ctggagcagc agcggcagcg ggagcagcag
     1381 cggcaggaag agctggagaa gcagcggctg gagcagcagc tgctcatcct gcggaacaag
     1441 gagaagagca aagagagtgc cattgccagc actgaggtaa agctgaggct ccaggaattc
     1501 ctcttgtcga agtcaaagga gcccacacca ggcggcctca accattccct cccacagcac
     1561 cccaaatgct ggggagccca ccatgcttct ttggaccaga gttcccctcc ccagagcggc
     1621 ccccctggga cgcctccctc ctacaaactg cctttgcctg ggccctacga cagtcgagac
     1681 gacttccccc tccgcaaaac agcctctgaa cccaacttga aagtgcgttc aaggctaaaa
     1741 cagaaggtgg ctgagcggag aagcagtccc ctcctgcgtc gcaaggatgg gactgttatt
     1801 agcaccttta agaagagagc tgttgagatc acaggtgccg ggcctggggc gtcgtccgtg
     1861 tgtaacagcg cacccggctc cggccccagc tctcccaaca gctcccacag caccatcgct
     1921 gagaatggct ttactggctc agtccccaac atccccactg agatgctccc tcagcaccga
     1981 gccctccctc tggacagctc ccccaaccag ttcagcctct acacgtctcc ttctctgccc
     2041 aacatctccc tagggctgca ggccacggtc actgtcacca actcacacct cactgcctcc
     2101 ccgaagctgt cgacacagca ggaggccgag aggcaggccc tccagtccct gcggcagggt
     2161 ggcacgctga ccggcaagtt catgagcaca tcctctattc ctggctgcct gctgggcgtg
     2221 gcactggagg gcgacgggag cccccacggg catgcctccc tgctgcagca tgtgctgttg
     2281 ctggagcagg cccggcagca gagcaccctc attgctgtgc cactccacgg gcagtcccca
     2341 ctagtgacgg gtgaacgtgt ggccaccagc atgcggacgg taggcaagct cccgcggcat
     2401 cggcccctga gccgcactca gtcctcaccg ctgccgcaga gtccccaggc cctgcagcag
     2461 ctggtcatgc aacaacagca ccagcagttc ctggagaagc agaagcagca gcagctacag
     2521 ctgggcaaga tcctcaccaa gacaggggag ctgcccaggc agcccaccac ccaccctgag
     2581 gagacagagg aggagctgac ggagcagcag gaggtcttgc tgggggaggg agccctgacc
     2641 atgccccggg agggctctac agagagtgag agcacacagg aagacctgga ggaggaggac
     2701 gaggaagacg atggggagga ggaggaggat tgcatccagg ttaaggacga ggagggcgag
     2761 agtggtgctg aggaggggcc cgacttggag gagcctggtg ctggatacaa aaaactgttc
     2821 tcagatgccc agccgctgca gcctttgcag gtgtaccagg cgcccctcag cctggccact
     2881 gtgccccacc aggccctggg ccgtacccag tcctcccctg ctgcccctgg gggcatgaag
     2941 agccccccag accagcccgt caagcacctc ttcaccacag gtgtggtcta cgacacgttc
     3001 atgctaaagc accagtgcat gtgcgggaac acacacgtgc accctgagca tgctggccgg
     3061 atccagagca tctggtcccg gctgcaggag acaggcctgc ttagcaagtg cgagcggatc
     3121 cgaggtcgca aagccacgct agatgagatc cagacagtgc actctgaata ccacaccctg
     3181 ctctatggga ccagtcccct caaccggcag aagctagaca gcaagaagtt gctcggcccc
     3241 atcagccaga agatgtatgc tgtgctgcct tgtgggggca tcggggtgga cagtgacacc
     3301 gtgtggaatg agatgcactc ctccagtgct gtgcgcatgg cagtgggctg cctgctggag
     3361 ctggccttca aggtggctgc aggagagctc aagaatggat ttgccatcat ccggccccca
     3421 ggacaccacg ccgaggaatc cacagccatg ggattctgct tcttcaactc tgtagccatc
     3481 accgcaaaac tcctacagca gaagttgaac gtgggcaagg tcctcatcgt ggactgggac
     3541 attcaccatg gcaatggcac ccagcaggcg ttctacaatg acccctctgt gctctacatc
     3601 tctctgcatc gctatgacaa cgggaacttc tttccaggct ctggggctcc tgaagaggtt
     3661 ggtggaggac caggcgtggg gtacaatgtg aacgtggcat ggacaggagg tgtggacccc
     3721 cccattggag acgtggagta ccttacagcc ttcaggacag tggtgatgcc cattgcccac
     3781 gagttctcac ctgatgtggt cctagtctcc gccgggtttg atgctgttga aggacatctg
     3841 tctcctctgg gtggctactc tgtcaccgcc agatgttttg gccacttgac caggcagctg
     3901 atgaccctgg cagggggccg ggtggtgctg gccctggagg gaggccatga cttgaccgcc
     3961 atctgtgatg cctctgaggc ttgtgtctcg gctctgctca gtgtagagct gcagcccttg
     4021 gatgaggcag tcttgcagca aaagcccaac atcaacgcag tggccacgct agagaaagtc
     4081 atcgagatcc agagcaaaca ctggagctgt gtgcagaagt tcgccgctgg tctgggccgg
     4141 tccctgcgag aggcccaagc aggtgagacc gaggaggccg agactgtgag cgccatggcc
     4201 ttgctgtcgg tgggggccga gcaggcccag gctgcggcag cccgggaaca cagccccagg
     4261 ccggcagagg agcccatgga gcaggagcct gccctgctcg agggaggaga ctacaaggac
     4321 gacgatgaca agtaatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     4381 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4441 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4501 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4561 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4621 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4681 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4741 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4801 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4861 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4921 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4981 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5041 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5101 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5161 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5221 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5281 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5341 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5401 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5461 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5521 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5581 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5641 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5701 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5761 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5821 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5881 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5941 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6001 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6061 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6121 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6181 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6241 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6301 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6361 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6421 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6481 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6541 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6601 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6661 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6721 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6781 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7441 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7681 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7741 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7801 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7861 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7921 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7981 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8041 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8101 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8161 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8221 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8281 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8341 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8401 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8461 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8521 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8581 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8641 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8701 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8761 agtgccacct gacg
//
