LOCUS       sequence-461730-        5108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KDAC8 (HDAC8) D101N in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5108)
  AUTHORS   Toro TB, Swanier JS, Bezue JA, Broussard CG, Watt TJ
  TITLE     Lysine Deacetylase Substrate Selectivity: A Dynamic Ionic 
            Interaction Specific to KDAC8.
  JOURNAL   Biochemistry. 2021 Aug 24;60(33):2524-2536. doi: 
            10.1021/acs.biochem.1c00384. Epub 2021 Aug 6.
  PUBMED    34357750
REFERENCE   2  (bases 1 to 5108)
  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: 
            "10.1021/acs.biochem.1c00384"; journalName: "Biochemistry"; date: 
            "2021-08-24- 24"; volume: "60"; issue: "33"; pages: "2524-2536"
COMMENT     Sequence Label: pJExpress401-KDAC8 D101N
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     575..594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     828..845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     943..960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     944..960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1416..1432
                     /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        join(1457..1471,1472..1477,1478..1494,1495..1501)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1477..1493
                     /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)."
     protein_bind    1507..1526
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             2713..2733
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2734..2751
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2956..2975)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3042..3133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3134..3943
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4026..5108
                     /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     complement(4045..4064)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc
       61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      241 tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      841 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
      901 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
      961 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
     1021 taaaaacccg cttcggcggg tttttttatg ggggggttta gggaaagagc atttgtcaga
     1081 atatttaagg gcgcctgtca ctttgcttga tatatgagaa ttatttaacc ttataaatga
     1141 gaaaaaagca acgcacttta aataagatac gttgcttttt cgattgatga acacctataa
     1201 ttaaactatt catctattat ttatgatttt ttgtatatac aatatttcta gtttgttaaa
     1261 gagaattaag aaaataaatc tcgaaaataa taaagggaaa atcagttttt gatatcaaaa
     1321 ttatacatgt caacgataat acaaaatata atacaaacta taagatgtta tcagtattta
     1381 ttatgcattt agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc
     1441 ttcatgcaca gtgaaatcat gaaaaattta tttgctttgt gagcggataa caattataat
     1501 atgtggaatt gtgagcgctc acaattccac aacggtttcc ctctagaaat aattttgttt
     1561 aacttttagg aggtaaaaca tatggaagag cctgaagagc ctgcagacag cggccaatcc
     1621 ctggtgccgg tttacattta cagcccggaa tatgtgagca tgtgcgatag cttggcgaaa
     1681 atccctaagc gcgcgagcat ggtccacagc ctgatcgagg cgtatgcgtt gcacaaacaa
     1741 atgcgtattg tgaaaccgaa agtggcatct atggaggaaa tggctacctt tcacacggac
     1801 gcgtacctgc aacatctgca gaaagtgagc caagaaggtg acgatgatca cccggacagc
     1861 attgagtatg gcctgggtta caattgtccg gccacggagg gtattttcga ctacgcggca
     1921 gcaatcggtg gtgcgacgat caccgcggca cagtgtctga tcgatggcat gtgcaaggtt
     1981 gccatcaatt ggagcggcgg ttggcatcac gcgaagaaag acgaagcatc cggtttctgc
     2041 tacctgaacg atgcggttct gggcatcctg cgtctgcgcc gtaagtttga acgcattctg
     2101 tacgttgacc tggatctgca tcacggtgat ggcgtcgagg acgccttctc ctttacgagc
     2161 aaagtcatga ccgtgtctct gcacaagttc agcccgggtt tctttccggg taccggcgat
     2221 gttagcgatg ttggtctggg taagggccgt tactacagcg tgaacgttcc gattcaggat
     2281 ggtatccagg acgagaaata ctatcagatc tgcgaaagcg tgctgaagga agtttaccag
     2341 gctttcaatc cgaaagccgt tgtcttgcag ctgggtgccg acacgatcgc aggtgacccg
     2401 atgtgcagct tcaatatgac cccggtgggt attggtaagt gtttgaaata cattctgcaa
     2461 tggcagctgg cgacgctgat tctgggtggc ggtggctaca acttggctaa caccgcgcgt
     2521 tgttggacct atttgaccgg tgtcatcctg ggcaagaccc tgagctcgga aattccggac
     2581 cacgagtttt tcaccgctta tggcccggat tatgtactgg agatcactcc gtcttgccgt
     2641 ccggaccgca acgagccaca ccgtattcaa caaattctga actatatcaa aggtaatctg
     2701 aagcacgtcg tcgagaatct gtattttcag ggccatcatc atcaccatca ctaactcgag
     2761 ccccaagggc gacaccccct aattagcccg ggcgaaaggc ccagtctttc gactgagcct
     2821 ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagtccc cacactacca
     2881 tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac caccgcgcta
     2941 ctgccgccag gcaaacaagg ggtgttatga gccatattca ggtataaatg ggctcgcgat
     3001 aatgttcaga attggttaat tggttgtaac actgacccct atttgtttat ttttctaaat
     3061 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3121 aaaaaggaag aatatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     3181 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3241 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3301 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3361 tatgccactt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3421 cactgcgatc cccggaaaaa cagcgttcca ggtattagaa gaatatcctg attcaggtga
     3481 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cactcgattc ctgtttgtaa
     3541 ttgtcctttt aacagcgatc gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa
     3601 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3661 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3721 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3781 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3841 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3901 gaataaattg caatttcatt tgatgctcga tgagtttttc taagcggcgc gccatcgaat
     3961 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     4021 tgaatatgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     4081 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     4141 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     4201 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     4261 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     4321 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     4381 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     4441 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     4501 ttattttctc ccatgaggac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     4561 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     4621 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     4681 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     4741 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     4801 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagata
     4861 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     4921 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     4981 tgccagtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     5041 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     5101 ggcagtga
//
