LOCUS       sequence-240596-        7555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses histone deacetylase 6 (HDAC6) catalytic domain 2 (CD2), 
            also known as zCD2, in Escherichia coli cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  AUTHORS   Hai Y, Christianson DW
  TITLE     Histone deacetylase 6 structure and molecular basis of catalysis and
            inhibition.
  JOURNAL   Nat Chem Biol. 2016 Sep;12(9):741-7. doi: 10.1038/nchembio.2134. 
            Epub 2016 Jul 25.
  PUBMED    27454933
REFERENCE   2  (bases 1 to 7555)
  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.2134"; journalName: "Nat Chem Biol"; date: 
            "2016-09"; volume: "12"; issue: "9"; pages: "741-7"
COMMENT     Sequence Label: zCD2
FEATURES             Location/Qualifiers
     source          1..7555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             308..325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             335..1435
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1409..1432
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1448..1468
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2627..2644
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2697..2715)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2711..2758
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2795..3250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2882..2901)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3092..3113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3343..4158)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3343..4158)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4066..4085
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4150..4169
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4280..4868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4769..4788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5022..5039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5054..5193
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5119..5138)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5279..5301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5295..5486)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6295..7377)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7339..7358
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7378..7455)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
      301 ttcttctcac catcaccatc accatggttc ttctatgaaa atcgaagaag gtaaactggt
      361 aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga
      421 gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc
      481 acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg
      541 tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa
      601 gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat
      661 cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac
      721 ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat
      781 gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc
      841 gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc
      901 gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac
      961 cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg
     1021 cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc
     1081 gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc
     1141 cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga
     1201 aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga
     1261 ggaagagttg gcgaaagatc cacgtattgc cgccactatg gaaaacgccc agaaaggtga
     1321 aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat
     1381 caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaatgg
     1441 gatcgaggaa aacctgtact tccaatccaa tgcaggtggc tctagtccga ttaccggtct
     1501 ggtttacgat cagcgtatga tgctgcatca caacatgtgg gactcgcatc acccggaact
     1561 gccgcaacgt atttcgcgca tctttagccg tcatgaagaa ctgcgcctgc tgtcacgttg
     1621 tcatcgcatt ccggcacgtc tggctaccga agaagaactg gcgctgtgtc attcctcaaa
     1681 acacatctcc attatcaaat cgagcgaaca catgaaaccg cgtgatctga atcgcctggg
     1741 cgacgaatac aactcaatct tcatctctaa cgaaagttat acctgtgcac tgctggccgc
     1801 gggttcttgt tttaacagtg cacaggctat tctgacgggc caagtgcgta atgcggtggc
     1861 catcgtgcgt ccgccgggcc atcacgcaga aaaagatacc gcttgcggct tctgcttttt
     1921 caacaccgca gctctgacgg cgcgttatgc ccagtctatt acccgtgaaa gtctgcgcgt
     1981 cctgatcgtg gattgggacg ttcatcacgg caatggtacg cagcatattt ttgaagaaga
     2041 tgactccgtc ctgtatatct cactgcaccg ttacgaagat ggtgcctttt tcccgaacag
     2101 cgaagatgca aattatgaca aagtcggcct gggtaaaggc cgcggttaca acgtgaatat
     2161 tccgtggaac ggcggtaaaa tgggcgatcc ggaatatatg gcggccttcc atcacctggt
     2221 gatgccgatc gcgcgtgaat ttgcgccgga actggtgctg gttagcgcgg gttttgatgc
     2281 agctcgcggt gacccgctgg gcggttttca ggtcaccccg gaaggttatg cacatctgac
     2341 gcaccaactg atgtctctgg ccgcgggtcg tgtgctgatt atcctggaag gcggttacaa
     2401 cctgaccagc atttcagaat cgatgagcat gtgtacgtct atgctgctgg gtgattcccc
     2461 gccgtcactg gaccatctga ccccgctgaa aacgagtgcg accgtgtcaa tcaataatgt
     2521 tctgcgtgct catgctccgt tttggtcttc actgcgttaa taacattgga agtggataac
     2581 ggatccgaat tcgagcgccg tcgacaagct tgcggccgca ctcgagcacc accaccacca
     2641 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2701 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2761 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2821 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2881 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2941 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3001 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3061 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3121 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3181 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa actagtaacg
     3241 tttacaattt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3301 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     3361 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     3421 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     3481 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     3541 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     3601 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     3661 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     3721 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     3781 cactgccagc gcatcaacaa tgttttcacc tgaatcagga tattcttcta atacctggaa
     3841 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     3901 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     3961 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     4021 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     4081 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     4141 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     4201 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4261 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4321 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4381 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4801 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4861 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4921 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4981 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5041 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5101 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     5161 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     5221 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5281 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     5341 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     5401 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     5461 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     5521 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     5581 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     5641 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     5701 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     5761 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     5821 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     5881 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     5941 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     6001 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     6061 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     6121 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     6181 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     6241 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     6301 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6361 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     6421 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6481 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6541 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     6601 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     6661 gggaacgata ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     6721 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     6781 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     6841 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     6901 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     6961 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     7021 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     7081 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     7141 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     7201 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     7261 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     7321 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     7381 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     7441 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     7501 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatg
//
