LOCUS       sequence-448043-        8350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KDAC6 (HDAC6) H611A in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8350)
  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 8350)
  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 H611A
FEATURES             Location/Qualifiers
     source          1..8350
                     /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="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             7651..7671
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             7672..7689
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7840..7974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7877..7896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7931..7950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8003..8168
                     /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 gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc ataccgtccc accatatgac ctcaaccggc
     4021 caggattcca ccacaaccag gcagcgaaga agtaggcaga acccccagtc gccccctcag
     4081 gactccagtg tcacttcgaa gcgaaatatt aaaaagggag ccgttccccg ctctatcccc
     4141 aatctagcgg aggtaaagaa gaaaggcaaa atgaagaagc tcggccaagc aatggaagaa
     4201 gacctaatcg tgggactgca agggatggat ctgaaccttg aggctgaagc actggctggc
     4261 actggcttgg tgttggatga gcagttaaat gaattccatt gcctctggga tgacagcttc
     4321 ccggaaggcc ctgagcggct ccatgccatc aaggagcaac tgatccagga gggcctccta
     4381 gatcgctgcg tgtcctttca ggcccggttt gctgaaaagg aagagctgat gttggttcac
     4441 agcctagaat atattgatct gatggaaaca acccagtaca tgaatgaggg agaactccgt
     4501 gtcctagcag acacctacga ctcagtttat ctgcatccga actcatactc ctgtgcctgc
     4561 ctggcctcag gctctgtcct caggctggtg gatgcggtcc tgggggctga gatccggaat
     4621 ggcatggcca tcattaggcc tcctggacat cacgcccagc acagtcttat ggatggctat
     4681 tgcatgttca accacgtggc tgtggcagcc cgctatgctc aacagaaaca ccgcatccgg
     4741 agggtcctta tcgtagattg ggatgtgcac cacggtcaag gaacacagtt caccttcgac
     4801 caggacccca gtgtcctcta tttctccatc caccgctacg agcagggtag gttctggccc
     4861 cacctgaagg cctctaactg gtccaccaca ggtttcggcc aaggccaagg atataccatc
     4921 aatgtgcctt ggaaccaggt ggggatgcgg gatgctgact acattgctgc tttcctgcac
     4981 gtcctgctgc cagtcgccct cgaattccag cctcagctgg tcctggtggc tgctggattt
     5041 gatgccctgc aaggggaccc caagggtgag atggccgcca ctccggcagg gttcgcccag
     5101 ctaacccacc tgctcatggg tctggcagga ggcaagctga tcctgtctct ggagggtggc
     5161 tacaacctcc gcgccctggc tgaaggcgtc agtgcttcgc tccacaccct tctgggagac
     5221 ccttgcccca tgctggagtc acctggtgcc ccctgccgga gtgcccaggc ttcagtttcc
     5281 tgtgctctgg aagcccttga gcccttctgg gaggttcttg tgagatcaac tgagaccgtg
     5341 gagagggaca acatggagga ggacaatgta gaggagagcg aggaggaagg accctgggag
     5401 ccccctgtgc tcccaatcct gacatggcca gtgctacagt ctcgcacagg gctggtctat
     5461 gaccaaaata tgatgaatca ctgcaacttg tgggacagcc accaccctga ggtaccccag
     5521 cgcatcttgc ggatcatgtg ccgtctggag gagctgggcc ttgccgggcg ctgcctcacc
     5581 ctgacaccgc gccctgccac agaggctgag ctgctcacct gtcacagtgc tgagtacgtg
     5641 ggtcatctcc gggccacaga gaaaatgaaa acccgggagc tgcaccgtga gagttccaac
     5701 tttgactcca tctatatctg ccccagtacc ttcgcctgtg cacagcttgc cactggcgct
     5761 gcctgccgcc tggtggaggc tgtgctctca ggagaggttc tgaatggtgc tgctgtggtg
     5821 cgtcccccag gacacgctgc agagcaggat gcagcttgcg gtttttgctt tttcaactct
     5881 gtggctgtgg ctgctcgcca tgcccagact atcagtgggc atgccctacg gatcctgatt
     5941 gtggattggg atgtccacca cggtaatgga actcagcaca tgtttgagga tgaccccagt
     6001 gtgctatatg tgtccctgca ccgctatgat catggcacct tcttccccat gggggatgag
     6061 ggtgccagca gccagatcgg ccgggctgcg ggcacaggct tcaccgtcaa cgtggcatgg
     6121 aacgggcccc gcatgggtga tgctgactac ctagctgcct ggcatcgcct ggtgcttccc
     6181 attgcctacg agtttaaccc agaactggtg ctggtctcag ctggctttga tgctgcacgg
     6241 ggggatccgc tggggggctg ccaggtgtca cctgagggtt atgcccacct cacccacctg
     6301 ctgatgggcc ttgccagtgg ccgcattatc cttatcctag agggtggcta taacctgaca
     6361 tccatctcag agtccatggc tgcctgcact cgctccctcc ttggagaccc accacccctg
     6421 ctgaccctgc cacggccccc actatcaggg gccctggcct caatcactga gaccatccaa
     6481 gtccatcgca gatactggcg cagcttacgg gtcatgaagg tagaagacag agaaggaccc
     6541 tccagttcta agttggtcac caagaaggca ccccaaccag ccaaacctag gttagctgag
     6601 cggatgacca cacgagaaaa gaaggttctg gaagcaggca tggggaaagt cacctcggca
     6661 tcatttgggg aagagtccac tccaggccag actaactcag agacagctgt ggtggccctc
     6721 actcaggacc agccctcaga ggcagccaca gggggagcca ctctggccca gaccatttct
     6781 gaggcagcca ttgggggagc catgctgggc cagaccacct cagaggaggc tgtcggggga
     6841 gccactccgg accagaccac ctcagaggag actgtgggag gagccattct ggaccagacc
     6901 acctcagagg atgctgttgg gggagccacg ctgggccaga ctacctcaga ggaggctgta
     6961 ggaggagcta cactggccca gaccacctcg gaggcagcca tggagggagc cacactggac
     7021 cagactacgt cagaggaggc tccagggggc accgagctga tccaaactcc tctagcctct
     7081 agcacagacc accagacccc cccaacctca cctgtgcagg gaactacacc ccagatatct
     7141 cccagtacac tgattgggag tctcaggacc ttggagctag gcagcgaatc tcagggggcc
     7201 tcagaatctc aggccccagg agaggagaac ctactaggag aggcagctgg aggtcaggac
     7261 atggctgatt cgatgctgat gcagggatct aggggcctca ctgatcaggc catattttat
     7321 gctgtgacac cactgccctg gtgtccccat ttggtggcag tatgccccat acctgcagca
     7381 ggcctagacg tgacccaacc ttgtggggac tgtggaacaa tccaagagaa ttgggtgtgt
     7441 ctctcttgct atcaggtcta ctgtggtcgt tacatcaatg gccacatgct ccaacaccat
     7501 ggaaattctg gacacccgct ggtcctcagc tacatcgacc tgtcagcctg gtgttactac
     7561 tgtcaggcct atgtccacca ccaggctctc ctagatgtga agaacatcgc ccaccagaac
     7621 aagtttgggg aggatatgcc ccacccacac gaaaatctgt atttccaagg tcaccatcac
     7681 caccatcact aactcgaggc atgcggtacc aagcttgtcg agaagtacta gaggatcata
     7741 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     7801 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     7861 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7921 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
     7981 ctgcttgagc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc
     8041 atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc
     8101 ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc
     8161 cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc
     8221 atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa atttttctgt
     8281 catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa
     8341 tggcgaatgg
//
