LOCUS       sequence-447914-        5247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KDAC7 (HDAC7) catalytic domain in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5247)
  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 5247)
  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-KDAC7 CD
FEATURES             Location/Qualifiers
     source          1..5247
                     /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             2852..2872
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2873..2890
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3095..3114)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        3181..3272
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3273..4082
                     /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             4165..5247
                     /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(4184..4203)
                     /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 atatgagccg cgcacagagc agcccagcag ccccagcatc
     1621 cctgagcgcg cctgagccgg cgagccaggc gcgtgttctg tctagctctg aaaccccggc
     1681 acgcaccctg ccgttcacca cgggcttgat ctacgattcc gtgatgctga agcaccagtg
     1741 ttcgtgtggc gacaacagcc gccacccgga gcatgctggt cgtatccaaa gcatttggtc
     1801 gcgcttgcaa gaacgcggcc tgcgtagcca gtgtgagtgc ctgcgtggtc gtaaagcaag
     1861 cctggaagaa ctgcaatcgg tccacagcga gcgtcatgtg ctgttgtatg gcaccaatcc
     1921 gctgagccgt ctgaagttgg ataatggcaa actggccggt ctgctggcac aacgtatgtt
     1981 tgtcatgctg ccgtgcggtg gtgttggtgt tgacaccgac acgatttgga atgagctgca
     2041 tagcagcaac gcggctcgct gggccgcagg ttctgtcacg gacctggctt ttaaggtggc
     2101 gagccgtgaa ctgaagaacg gctttgccgt tgtgcgtccg ccaggtcacc acgccgatca
     2161 ttccacggcg atgggctttt gctttttcaa cagcgtagcg atcgcgtgtc gtcaactgca
     2221 gcagcagagc aaagcgtcca agattttgat cgtggactgg gatgtccatc atggtaatgg
     2281 tacccaacag actttctacc aggatccgag cgttttgtat atctctctgc accgtcacga
     2341 cgatggcaac ttctttccgg gttccggtgc ggttgatgaa gttggtgcag gcagcggcga
     2401 aggtttcaac gtgaacgttg cgtgggccgg tggcctggac ccgccgatgg gtgacccgga
     2461 gtacctggcg gcattccgta ttgttgtgat gccgatcgcg cgtgagttca gcccggatct
     2521 ggttctggtc agcgcgggtt tcgatgctgc cgagggtcac ccggcaccgc tgggtggcta
     2581 tcacgtgtcc gcgaagtgct ttggctatat gacccagcaa ctgatgaact tggcgggtgg
     2641 tgccgtcgtc ctggcgttgg aaggtggtca tgacctgacc gcgatttgcg acgcgagcga
     2701 ggcgtgcgtg gcagctctgc tgggcaatcg tgttgacccg ctgagcgagg aaggttggaa
     2761 acaaaaacct aatctgaatg ccatacgtag cctggaggca gtgattcgcg tccatagcaa
     2821 atactggggt tgtatgcaac gcctggcgag cgagaatctg tacttccaag gccatcacca
     2881 tcaccaccac taactcgagc cccaagggcg acacccccta attagcccgg gcgaaaggcc
     2941 cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg
     3001 gggagtcccc acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc
     3061 aggtgggacc accgcgctac tgccgccagg caaacaaggg gtgttatgag ccatattcag
     3121 gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta
     3181 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3241 aaatgcttca ataatattga aaaaggaaga atatgagcca tattcaacgg gaaacgtcga
     3301 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     3361 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     3421 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     3481 taaactggct gacggaattt atgccacttc cgaccatcaa gcattttatc cgtactcctg
     3541 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcgttccag gtattagaag
     3601 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     3661 actcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc ctcgctcagg
     3721 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3781 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3841 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3901 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3961 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     4021 gtattgataa tcctgatatg aataaattgc aatttcattt gatgctcgat gagtttttct
     4081 aagcggcgcg ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     4141 gagagtcaat tcagggtggt gaatatgaaa ccagtaacgt tatacgatgt cgcagagtat
     4201 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     4261 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     4321 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     4381 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     4441 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     4501 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     4561 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     4621 cagacaccca tcaacagtat tattttctcc catgaggacg gtacgcgact gggcgtggag
     4681 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     4741 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     4801 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     4861 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     4921 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     4981 tacgacgata ccgaagatag ctcatgttat atcccgccgt taaccaccat caaacaggat
     5041 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     5101 gtgaagggca atcagctgtt gccagtctca ctggtgaaaa gaaaaaccac cctggcgccc
     5161 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5221 gtttcccgac tggaaagcgg gcagtga
//
