LOCUS       sequence-215891-        9028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PYL-HDAC5 H1006Y expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9028)
  AUTHORS   Yang B, Borgeaud AC, Buricova M, Aeschbach L, Rodriguez-Lima O, Ruiz
            Buendia GA, Cinesi C, Taylor AS, Baubec T, Dion V
  TITLE     Expanded CAG/CTG repeats resist gene silencing mediated by targeted 
            Epigenome editing.
  JOURNAL   Hum Mol Genet. 2021 Sep 7. pii: 6365982. doi: 10.1093/hmg/ddab255.
  PUBMED    34494094
REFERENCE   2  (bases 1 to 9028)
  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: "Hum Mol 
            Genet. 2021 Sep 7. pii: 6365982. doi: 10.1093/hmg/ddab255."
COMMENT     Sequence Label: pAB-HDAC5(mut-H-Y)
FEATURES             Location/Qualifiers
     source          1..9028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      935..944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1478..1549
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1553..1573
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1574..1598
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4970..4994)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5035..5052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5041..5265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5311..5739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5398..5417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5608..5629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5748..5768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5753..6083
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5934..6069
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5996..6015
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6131..6178
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6197..6571
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6701..6822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6738..6757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6792..6811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6871..6887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6871..6887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6884..6906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6895..6911
                     /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        complement(join(6919..6925,6926..6943,6944..6949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6964..6985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7102..7119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7273..7861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7353..7372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8032..8823,8824..8892))
                     /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     8655..8674
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8893..8997)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggccaccc aggacgagtt
      961 cacccagctg agccagtcta tcgccgagtt ccacacctac cagctgggca acggccggtg
     1021 cagcagcctg ctggcccaga gaatccacgc ccctcccgag acagtgtggt ccgtcgtgcg
     1081 gagattcgac cggccccaga tctacaagca cttcatcaag agctgcaacg tgtccgagga
     1141 cttcgagatg agagtgggct gcacccggga cgtgaacgtg atcagcggcc tgcccgccaa
     1201 caccagcaga gagagactgg acctgctgga cgacgacaga agagtgaccg gcttcagcat
     1261 caccggcgga gagcaccggc tgcggaacta caagagcgtg accaccgtgc acagattcga
     1321 gaaagaggaa gaagaggaac ggatctggac cgtggtgctg gaaagctacg tggtggacgt
     1381 gcccgagggc aacagcgaag aggacacccg gctgttcgcc gacaccgtga tccggctgaa
     1441 cctgcagaag ctggcctcca tcaccgaggc catgaacgac tacaaggacg acgacgacaa
     1501 ggactataag gacgatgatg ataaggatta caaagatgac gacgataagc cccccaagaa
     1561 gaagcggaaa gtcacaagtt tgtacaaaaa agcaggctca atgaactctc ccaacgagtc
     1621 ggatgggatg tcaggtcggg aaccatcctt ggaaatcctg ccgcggactt ctctgcacag
     1681 catccctgtg acagtggagg tgaagccggt gctgccaaga gccatgccca gttccatggg
     1741 gggtgggggt ggaggcagcc ccagccctgt ggagctacgg ggggctctgg tgggctctgt
     1801 ggaccccaca ctgcgggagc agcaactgca gcaggagctc ctggcgctca agcagcagca
     1861 gcagctgcag aagcagctcc tgttcgctga gttccagaaa cagcatgacc acctgacaag
     1921 gcagcatgag gtccagctgc agaagcacct caagcagcag caggagatgc tggcagccaa
     1981 gcagcagcag gagatgctgg cagccaagcg gcagcaggag ctggagcagc agcggcagcg
     2041 ggagcagcag cggcaggaag agctggagaa gcagcggctg gagcagcagc tgctcatcct
     2101 gcggaacaag gagaagagca aagagagtgc cattgccagc actgaggtaa agctgaggct
     2161 ccaggaattc ctcttgtcga agtcaaagga gcccacacca ggcggcctca accattccct
     2221 cccacagcac cccaaatgct ggggagccca ccatgcttct ttggaccaga gttcccctcc
     2281 ccagagcggc ccccctggga cgcctccctc ctacaaactg cctttgcctg ggccctacga
     2341 cagtcgagac gacttccccc tccgcaaaac agcctctgaa cccaacttga aagtgcgttc
     2401 aaggctaaaa cagaaggtgg ctgagcggag aagcagtccc ctcctgcgtc gcaaggatgg
     2461 gactgttatt agcaccttta agaagagagc tgttgagatc acaggtgccg ggcctggggc
     2521 gtcgtccgtg tgtaacagcg cacccggctc cggccccagc tctcccaaca gctcccacag
     2581 caccatcgct gagaatggct ttactggctc agtccccaac atccccactg agatgctccc
     2641 tcagcaccga gccctccctc tggacagctc ccccaaccag ttcagcctct acacgtctcc
     2701 ttctctgccc aacatctccc tagggctgca ggccacggtc actgtcacca actcacacct
     2761 cactgcctcc ccgaagctgt cgacacagca ggaggccgag aggcaggccc tccagtccct
     2821 gcggcagggt ggcacgctga ccggcaagtt catgagcaca tcctctattc ctggctgcct
     2881 gctgggcgtg gcactggagg gcgacgggag cccccacggg catgcctccc tgctgcagca
     2941 tgtgctgttg ctggagcagg cccggcagca gagcaccctc attgctgtgc cactccacgg
     3001 gcagtcccca ctagtgacgg gtgaacgtgt ggccaccagc atgcggacgg taggcaagct
     3061 cccgcggcat cggcccctga gccgcactca gtcctcaccg ctgccgcaga gtccccaggc
     3121 cctgcagcag ctggtcatgc aacaacagca ccagcagttc ctggagaagc agaagcagca
     3181 gcagctacag ctgggcaaga tcctcaccaa gacaggggag ctgcccaggc agcccaccac
     3241 ccaccctgag gagacagagg aggagctgac ggagcagcag gaggtcttgc tgggggaggg
     3301 agccctgacc atgccccggg agggctccac agagagtgag agcacacagg aagacctgga
     3361 ggaggaggac gaggaagagg atggggagga ggaggaggat tgcatccagg ttaaggacga
     3421 ggagggcgag agtggtgctg aggaggggcc cgacttggag gagcctggtg ctggatacaa
     3481 aaaactgttc tcagatgccc agccgctgca gcctttgcag gtgtaccagg cgcccctcag
     3541 cctggccact gtgccccacc aggccctggg ccgtacccag tcctcccctg ctgcccctgg
     3601 gggcatgaag agccccccag accagcccgt caagcacctc ttcaccacag gtgtggtcta
     3661 cgacacgttc atgctaaagc accagtgcat gtgcgggaac acacacgtgc accctgagca
     3721 tgctggccgg atccagagca tctggtcccg gctgcaggag acaggcctgc ttagcaagtg
     3781 cgagcggatc cgaggtcgca aagccacgct agatgagatc cagacagtgc actctgaata
     3841 ccacaccctg ctctatggga ccagtcccct caaccggcag aagctagaca gcaagaagtt
     3901 gctcggcccc atcagccaga agatgtatgc tgtgctgcct tgtgggggca tcggggtgga
     3961 cagtgacacc gtgtggaatg agatgcactc ctccagtgct gtgcgcatgg cagtgggctg
     4021 cctgctggag ctggccttca aggtggctgc aggagagctc aagaatggat ttgccatcat
     4081 ccggccccca ggacaccacg ccgaggaatc cacagccatg ggattctgct tcttcaactc
     4141 tgtagccatc accgcaaaac tcctacagca gaagttgaac gtgggcaagg tcctcatcgt
     4201 ggactgggac attcaccatg gcaatggcac ccagcaggcg ttctacaatg acccctctgt
     4261 gctctacatc tctctgcatc gctatgacaa cgggaacttc tttccaggct ctggggctcc
     4321 tgaagaggtt ggtggaggac caggcgtggg gtacaatgtg aacgtggcat ggacaggagg
     4381 tgtggacccc cccattggag acgtggagta ccttacagcc ttcaggacag tggtgatgcc
     4441 cattgcccac gagttctcac ctgatgtggt cctagtctcc gccgggtttg atgctgttga
     4501 aggacatctg tctcctctgg gtggctactc tgtcaccgcc agatgttttg gccacttgac
     4561 caggcagctg atgaccctgg cagggggccg ggtggtgctg gccctggagg gaggctatga
     4621 cttgaccgcc atctgtgatg cctctgaggc ttgtgtctcg gctctgctca gtgtagagct
     4681 gcagcccttg gatgaggcag tcttgcagca aaagcccaac atcaacgcag tggccacgct
     4741 agagaaagtc atcgagatcc agagcaaaca ctggagctgt gtgcagaagt tcgccgctgg
     4801 tctgggccgg tccctgcgag aggcccaagc aggtgagacc gaggaggccg agactgtgag
     4861 cgccatggcc ttgctgtcgg tgggggccga gcaggcccag gctgcggcag cccgggaaca
     4921 cagccccagg ccggcagagg agcccatgga gcaggagcct gccctgtgaa cccagctttc
     4981 ttgtacaaag tggtcatccg cggtctagag ggcccgttta aacccgctga tcagcctcga
     5041 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5101 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5161 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5221 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     5281 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     5341 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5401 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5461 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5521 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5581 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5641 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     5701 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     5761 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     5821 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5881 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5941 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6001 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6061 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6121 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     6181 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     6241 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     6301 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     6361 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     6421 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     6481 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     6541 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     6601 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6661 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6721 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6781 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6841 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6901 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6961 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7021 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7081 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7141 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7201 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7261 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7321 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7381 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7441 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     7501 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7561 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7621 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7681 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7741 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7801 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7861 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7921 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7981 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8041 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8101 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8161 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8221 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8281 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8341 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8401 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8461 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8521 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8581 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8641 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8701 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8761 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8821 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8881 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8941 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9001 atttccccga aaagtgccac ctgacgtc
//
