LOCUS       sequence-447933-        5874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KDAC8 (HDAC8) H143A in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5874)
  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 5874)
  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: pFastBacI-KDAC8 H143A
FEATURES             Location/Qualifiers
     source          1..5874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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     complement(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3543..3548,3549..3565,3566..3571))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3923..4014
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3939..3956
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5175..5195
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5196..5213
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5364..5498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5401..5420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5455..5474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5527..5692
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctgcgta acatcgttgc
     3361 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     3421 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     3481 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     3541 acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg
     3601 gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg
     3661 cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct
     3721 tctacggcaa ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc
     3781 cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc
     3841 tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac
     3901 gtatactccg gaatattaat agatcatgga gataattaaa atgataacca tctcgcaaat
     3961 aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat aaatattccg
     4021 gattattcat accgtcccac catatggaag agcctgaaga gcctgcagac agcggccaat
     4081 ccctggtgcc ggtttacatt tacagcccgg aatatgtgag catgtgcgat agcttggcga
     4141 aaatccctaa gcgcgcgagc atggtccaca gcctgatcga ggcgtatgcg ttgcacaaac
     4201 aaatgcgtat tgtgaaaccg aaagtggcat ctatggagga aatggctacc tttcacacgg
     4261 acgcgtacct gcaacatctg cagaaagtga gccaagaagg tgacgatgat cacccggaca
     4321 gcattgagta tggcctgggt tacgattgtc cggccacgga gggtattttc gactacgcgg
     4381 cagcaatcgg tggtgcgacg atcaccgcgg cacagtgtct gatcgatggc atgtgcaagg
     4441 ttgccatcaa ttggagcggc ggttggcatg cagcgaagaa agacgaagca tccggtttct
     4501 gctacctgaa cgatgcggtt ctgggcatcc tgcgtctgcg ccgtaagttt gaacgcattc
     4561 tgtacgttga cctggatctg catcacggtg atggcgtcga ggacgccttc tcctttacga
     4621 gcaaagtcat gaccgtgtct ctgcacaagt tcagcccggg tttctttccg ggtaccggcg
     4681 atgttagcga tgttggtctg ggtaagggcc gttactacag cgtgaacgtt ccgattcagg
     4741 atggtatcca ggacgagaaa tactatcaga tctgcgaaag cgtgctgaag gaagtttacc
     4801 aggctttcaa tccgaaagcc gttgtcttgc agctgggtgc cgacacgatc gcaggtgacc
     4861 cgatgtgcag cttcaatatg accccggtgg gtattggtaa gtgtttgaaa tacattctgc
     4921 aatggcagct ggcgacgctg attctgggtg gcggtggcta caacttggct aacaccgcgc
     4981 gttgttggac ctatttgacc ggtgtcatcc tgggcaagac cctgagctcg gaaattccgg
     5041 accacgagtt tttcaccgct tatggcccgg attatgtact ggagatcact ccgtcttgcc
     5101 gtccggaccg caacgagcca caccgtattc aacaaattct gaactatatc aaaggtaatc
     5161 tgaagcacgt cgtcgagaat ctgtattttc agggccatca tcatcaccat cactaactcg
     5221 aggcatgcgg taccaagctt gtcgagaagt actagaggat cataatcagc cataccacat
     5281 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5341 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5401 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5461 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     5521 gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     5581 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     5641 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     5701 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     5761 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     5821 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga atgg
//
