LOCUS       sequence-447911-        5292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KDAC4 (HDAC4) catalytic domain in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5292)
  AUTHORS   Toro TB, Painter RG, Haynes RA, Glotser EY, Bratton MR, Bryant JR, 
            Nichols KA, Matthew-Onabanjo AN, Matthew AN, Bratcher DR, Perry CD, 
            Watt TJ
  TITLE     Purification of metal-dependent lysine deacetylases with 
            consistently high activity.
  JOURNAL   Protein Expr Purif. 2018 Jan;141:1-6. doi: 
            10.1016/j.pep.2017.08.009. Epub 2017 Aug 24.
  PUBMED    28843507
REFERENCE   2  (bases 1 to 5292)
  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: "Protein 
            Expr Purif."; date: "2018-01"; volume: "141:1-6. doi"; pages: " 
            10.1016/j.pep.2017.08.009. Epub 2017 Aug 24"
COMMENT     Sequence Label: pJExpress401-KDAC4 CD
FEATURES             Location/Qualifiers
     source          1..5292
                     /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    1417..1433
                     /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(1458..1472,1473..1478,1479..1495,1496..1502)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1478..1494
                     /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    1508..1527
                     /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             2897..2917
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2918..2935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3140..3159)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3226..3317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3318..4127
                     /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             4210..5292
                     /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(4229..4248)
                     /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 gggggagttt agggaaagag catttgtcag
     1081 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
     1141 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
     1201 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
     1261 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
     1321 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
     1381 attatgcatt tagaataaat tttgtgtcgc ccttaattgt gagcggataa caattacgag
     1441 cttcatgcac agtgaaatca tgaaaaattt atttgctttg tgagcggata acaattataa
     1501 tatgtggaat tgtgagcgct cacaattcca caacggtttc cctctagaaa taattttgtt
     1561 taacttttag gaggtaaaac atatgacgaa accgcgcttc accaccggcc tggtctacga
     1621 caccctgatg ctgaagcacc aatgcacatg tggcagcagc agctctcacc cggagcacgc
     1681 aggccgtatc caatccattt ggtcccgcct gcaggaaacg ggcctgcgcg gcaagtgcga
     1741 atgcattcgt ggccgtaaag ctaccctgga agagttgcag actgtgcact ccgaggcaca
     1801 caccttgttg tatggtacga atccgctgaa ccgccagaaa ctggactcca agaaactgtt
     1861 gggcagcctg gctagcgttt tcgtgcgcct gccgtgtggt ggtgttggcg tcgactctga
     1921 cacgatctgg aacgaggtcc atagcgcagg tgcggcacgt ctggcagtgg gctgcgttgt
     1981 ggagctggtg tttaaggttg cgaccggtga gctgaagaat ggttttgcgg tggtgcgtcc
     2041 gccaggtcac catgctgaag aaagcacccc gatgggtttc tgttacttta acagcgtggc
     2101 cgttgcggcc aaactgctgc aacaacgtct gagcgtcagc aaaatcctga tcgtcgactg
     2161 ggatgtccat cacggtaacg gtacgcagca ggccttctat tctgatccga gcgtcctgta
     2221 tatgagcctg caccgttacg acgacggcaa tttctttccg ggcagcggtg cgccggatga
     2281 agttggtacc ggtccgggtg tgggctttaa tgtgaacatg gcctttaccg gtggtctgga
     2341 ccctccgatg ggtgatgcag aatacctggc ggcgttccgc acggttgtaa tgccgatcgc
     2401 gagcgagttc gcaccggatg ttgtcctggt tagctctggc ttcgacgcgg tggagggtca
     2461 tccgacgcct ctgggtggtt acaacctgag cgcgcgttgt tttggttatc tgaccaagca
     2521 actgatgggt ttggcgggtg gtcgtatcgt gctggcgctg gagggtggtc atgatctgac
     2581 cgcgatttgc gatgctagcg aggcgtgtgt gagcgccctg ctgggcaatg agctggatcc
     2641 gttgccggaa aaagttctgc agcagcgtcc gaatgcgaac gccgttcgtt cgatggagaa
     2701 agttatggag attcatagca agtactggcg ttgtttgcag cgcactacga gcaccgctgg
     2761 ccgtagcctg attgaggcgc aaacctgcga aaacgaggaa gccgaaaccg ttacggcaat
     2821 ggcaagcttg agcgtcggcg tcaaaccagc ggagaagcgc cctgacgaag agccgatgga
     2881 ggaagaaccg ccgctggaaa atctgtattt ccaaggtcat catcaccacc accattaact
     2941 cgagccccaa gggcgacacc ccctaattag cccgggcgaa aggcccagtc tttcgactga
     3001 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcatggggag tccccacact
     3061 accatcggcg ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc
     3121 gctactgccg ccaggcaaac aaggggtgtt atgagccata ttcaggtata aatgggctcg
     3181 cgataatgtt cagaattggt taattggttg taacactgac ccctatttgt ttatttttct
     3241 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3301 attgaaaaag gaagaatatg agccatattc aacgggaaac gtcgaggccg cgattaaatt
     3361 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     3421 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     3481 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     3541 aatttatgcc acttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     3601 tcaccactgc gatccccgga aaaacagcgt tccaggtatt agaagaatat cctgattcag
     3661 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcactcg attcctgttt
     3721 gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga
     3781 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     3841 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg
     3901 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     3961 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     4021 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     4081 atatgaataa attgcaattt catttgatgc tcgatgagtt tttctaagcg gcgcgccatc
     4141 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     4201 gtggtgaata tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     4261 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     4321 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     4381 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     4441 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     4501 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     4561 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     4621 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     4681 agtattattt tctcccatga ggacggtacg cgactgggcg tggagcatct ggtcgcattg
     4741 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     4801 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     4861 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     4921 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     4981 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     5041 gatagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     5101 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     5161 ctgttgccag tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     5221 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5281 agcgggcagt ga
//
