LOCUS       sequence-448041-        8369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KDAC6 (HDAC6) H216A in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8369)
  AUTHORS   Toro TB, Bornes KE, Watt TJ
  TITLE     Lysine Deacetylase Substrate Selectivity: Distinct Interaction 
            Surfaces Drive Positive and Negative Selection for Residues 
            Following Acetyllysine.
  JOURNAL   Biochemistry. 2023 May 2;62(9):1464-1483. doi: 
            10.1021/acs.biochem.3c00001. Epub 2023 Apr 12.
  PUBMED    37043688
REFERENCE   2  (bases 1 to 8369)
  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.3c00001"; journalName: "Biochemistry"; date: 
            "2023-05-2- 2"; volume: "62"; issue: "9"; pages: "1464-1483"
COMMENT     Sequence Label: pFastBacI-KDAC6 H216A
FEATURES             Location/Qualifiers
     source          1..8369
                     /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(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3904..3995
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3920..3937
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             7670..7690
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7691..7708
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7859..7993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7896..7915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7950..7969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8022..8187
                     /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 ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     3901 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt
     3961 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca
     4021 ccatatgacc tcaaccggcc aggattccac cacaaccagg cagcgaagaa gtaggcagaa
     4081 cccccagtcg ccccctcagg actccagtgt cacttcgaag cgaaatatta aaaagggagc
     4141 cgttccccgc tctatcccca atctagcgga ggtaaagaag aaaggcaaaa tgaagaagct
     4201 cggccaagca atggaagaag acctaatcgt gggactgcaa gggatggatc tgaaccttga
     4261 ggctgaagca ctggctggca ctggcttggt gttggatgag cagttaaatg aattccattg
     4321 cctctgggat gacagcttcc cggaaggccc tgagcggctc catgccatca aggagcaact
     4381 gatccaggag ggcctcctag atcgctgcgt gtcctttcag gcccggtttg ctgaaaagga
     4441 agagctgatg ttggttcaca gcctagaata tattgatctg atggaaacaa cccagtacat
     4501 gaatgaggga gaactccgtg tcctagcaga cacctacgac tcagtttatc tgcatccgaa
     4561 ctcatactcc tgtgcctgcc tggcctcagg ctctgtcctc aggctggtgg atgcggtcct
     4621 gggggctgag atccggaatg gcatggccat cattaggcct cctggacatg cagcccagca
     4681 cagtcttatg gatggctatt gcatgttcaa ccacgtggct gtggcagccc gctatgctca
     4741 acagaaacac cgcatccgga gggtccttat cgtagattgg gatgtgcacc acggtcaagg
     4801 aacacagttc accttcgacc aggaccccag tgtcctctat ttctccatcc accgctacga
     4861 gcagggtagg ttctggcccc acctgaaggc ctctaactgg tccaccacag gtttcggcca
     4921 aggccaagga tataccatca atgtgccttg gaaccaggtg gggatgcggg atgctgacta
     4981 cattgctgct ttcctgcacg tcctgctgcc agtcgccctc gaattccagc ctcagctggt
     5041 cctggtggct gctggatttg atgccctgca aggggacccc aagggtgaga tggccgccac
     5101 tccggcaggg ttcgcccagc taacccacct gctcatgggt ctggcaggag gcaagctgat
     5161 cctgtctctg gagggtggct acaacctccg cgccctggct gaaggcgtca gtgcttcgct
     5221 ccacaccctt ctgggagacc cttgccccat gctggagtca cctggtgccc cctgccggag
     5281 tgcccaggct tcagtttcct gtgctctgga agcccttgag cccttctggg aggttcttgt
     5341 gagatcaact gagaccgtgg agagggacaa catggaggag gacaatgtag aggagagcga
     5401 ggaggaagga ccctgggagc cccctgtgct cccaatcctg acatggccag tgctacagtc
     5461 tcgcacaggg ctggtctatg accaaaatat gatgaatcac tgcaacttgt gggacagcca
     5521 ccaccctgag gtaccccagc gcatcttgcg gatcatgtgc cgtctggagg agctgggcct
     5581 tgccgggcgc tgcctcaccc tgacaccgcg ccctgccaca gaggctgagc tgctcacctg
     5641 tcacagtgct gagtacgtgg gtcatctccg ggccacagag aaaatgaaaa cccgggagct
     5701 gcaccgtgag agttccaact ttgactccat ctatatctgc cccagtacct tcgcctgtgc
     5761 acagcttgcc actggcgctg cctgccgcct ggtggaggct gtgctctcag gagaggttct
     5821 gaatggtgct gctgtggtgc gtcccccagg acaccacgca gagcaggatg cagcttgcgg
     5881 tttttgcttt ttcaactctg tggctgtggc tgctcgccat gcccagacta tcagtgggca
     5941 tgccctacgg atcctgattg tggattggga tgtccaccac ggtaatggaa ctcagcacat
     6001 gtttgaggat gaccccagtg tgctatatgt gtccctgcac cgctatgatc atggcacctt
     6061 cttccccatg ggggatgagg gtgccagcag ccagatcggc cgggctgcgg gcacaggctt
     6121 caccgtcaac gtggcatgga acgggccccg catgggtgat gctgactacc tagctgcctg
     6181 gcatcgcctg gtgcttccca ttgcctacga gtttaaccca gaactggtgc tggtctcagc
     6241 tggctttgat gctgcacggg gggatccgct ggggggctgc caggtgtcac ctgagggtta
     6301 tgcccacctc acccacctgc tgatgggcct tgccagtggc cgcattatcc ttatcctaga
     6361 gggtggctat aacctgacat ccatctcaga gtccatggct gcctgcactc gctccctcct
     6421 tggagaccca ccacccctgc tgaccctgcc acggccccca ctatcagggg ccctggcctc
     6481 aatcactgag accatccaag tccatcgcag atactggcgc agcttacggg tcatgaaggt
     6541 agaagacaga gaaggaccct ccagttctaa gttggtcacc aagaaggcac cccaaccagc
     6601 caaacctagg ttagctgagc ggatgaccac acgagaaaag aaggttctgg aagcaggcat
     6661 ggggaaagtc acctcggcat catttgggga agagtccact ccaggccaga ctaactcaga
     6721 gacagctgtg gtggccctca ctcaggacca gccctcagag gcagccacag ggggagccac
     6781 tctggcccag accatttctg aggcagccat tgggggagcc atgctgggcc agaccacctc
     6841 agaggaggct gtcgggggag ccactccgga ccagaccacc tcagaggaga ctgtgggagg
     6901 agccattctg gaccagacca cctcagagga tgctgttggg ggagccacgc tgggccagac
     6961 tacctcagag gaggctgtag gaggagctac actggcccag accacctcgg aggcagccat
     7021 ggagggagcc acactggacc agactacgtc agaggaggct ccagggggca ccgagctgat
     7081 ccaaactcct ctagcctcta gcacagacca ccagaccccc ccaacctcac ctgtgcaggg
     7141 aactacaccc cagatatctc ccagtacact gattgggagt ctcaggacct tggagctagg
     7201 cagcgaatct cagggggcct cagaatctca ggccccagga gaggagaacc tactaggaga
     7261 ggcagctgga ggtcaggaca tggctgattc gatgctgatg cagggatcta ggggcctcac
     7321 tgatcaggcc atattttatg ctgtgacacc actgccctgg tgtccccatt tggtggcagt
     7381 atgccccata cctgcagcag gcctagacgt gacccaacct tgtggggact gtggaacaat
     7441 ccaagagaat tgggtgtgtc tctcttgcta tcaggtctac tgtggtcgtt acatcaatgg
     7501 ccacatgctc caacaccatg gaaattctgg acacccgctg gtcctcagct acatcgacct
     7561 gtcagcctgg tgttactact gtcaggccta tgtccaccac caggctctcc tagatgtgaa
     7621 gaacatcgcc caccagaaca agtttgggga ggatatgccc cacccacacg aaaatctgta
     7681 tttccaaggt caccatcacc accatcacta actcgaggca tgcggtacca agcttgtcga
     7741 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     7801 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     7861 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     7921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     7981 tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata agtgaaatct
     8041 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     8101 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     8161 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat
     8221 caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac
     8281 agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     8341 cgcttatttg cagcctgaat ggcgaatgg
//
