LOCUS       sequence-225199-        7267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PYL-HDAC5 H1006A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7267)
  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 7267)
  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-A)
FEATURES             Location/Qualifiers
     source          1..7267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..23)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        195..574
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        575..778
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     729..749
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        823..841
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1438..1509
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1513..1533
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1534..1558
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4930..5029)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(5047..5065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5070..5086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5070..5086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5083..5105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5094..5110
                     /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(5118..5124,5125..5142,5143..5148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5163..5184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5301..5318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5472..6060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5552..5571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6231..7022,7023..7091))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNIGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6854..6873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7092..7196)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
       61 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
      121 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      181 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      241 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      301 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      361 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      421 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      481 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      541 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      601 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      661 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      721 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      781 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      841 ggagacccaa gctggctagc gtttaaactt aagcttggta ccgagctcgg atccgccacc
      901 atggccaccc aggacgagtt cacccagctg agccagtcta tcgccgagtt ccacacctac
      961 cagctgggca acggccggtg cagcagcctg ctggcccaga gaatccacgc ccctcccgag
     1021 acagtgtggt ccgtcgtgcg gagattcgac cggccccaga tctacaagca cttcatcaag
     1081 agctgcaacg tgtccgagga cttcgagatg agagtgggct gcacccggga cgtgaacgtg
     1141 atcagcggcc tgcccgccaa caccagcaga gagagactgg acctgctgga cgacgacaga
     1201 agagtgaccg gcttcagcat caccggcgga gagcaccggc tgcggaacta caagagcgtg
     1261 accaccgtgc acagattcga gaaagaggaa gaagaggaac ggatctggac cgtggtgctg
     1321 gaaagctacg tggtggacgt gcccgagggc aacagcgaag aggacacccg gctgttcgcc
     1381 gacaccgtga tccggctgaa cctgcagaag ctggcctcca tcaccgaggc catgaacgac
     1441 tacaaggacg acgacgacaa ggactataag gacgatgatg ataaggatta caaagatgac
     1501 gacgataagc cccccaagaa gaagcggaaa gtcacaagtt tgtacaaaaa agcaggctca
     1561 atgaactctc ccaacgagtc ggatgggatg tcaggtcggg aaccatcctt ggaaatcctg
     1621 ccgcggactt ctctgcacag catccctgtg acagtggagg tgaagccggt gctgccaaga
     1681 gccatgccca gttccatggg gggtgggggt ggaggcagcc ccagccctgt ggagctacgg
     1741 ggggctctgg tgggctctgt ggaccccaca ctgcgggagc agcaactgca gcaggagctc
     1801 ctggcgctca agcagcagca gcagctgcag aagcagctcc tgttcgctga gttccagaaa
     1861 cagcatgacc acctgacaag gcagcatgag gtccagctgc agaagcacct caagcagcag
     1921 caggagatgc tggcagccaa gcagcagcag gagatgctgg cagccaagcg gcagcaggag
     1981 ctggagcagc agcggcagcg ggagcagcag cggcaggaag agctggagaa gcagcggctg
     2041 gagcagcagc tgctcatcct gcggaacaag gagaagagca aagagagtgc cattgccagc
     2101 actgaggtaa agctgaggct ccaggaattc ctcttgtcga agtcaaagga gcccacacca
     2161 ggcggcctca accattccct cccacagcac cccaaatgct ggggagccca ccatgcttct
     2221 ttggaccaga gttcccctcc ccagagcggc ccccctggga cgcctccctc ctacaaactg
     2281 cctttgcctg ggccctacga cagtcgagac gacttccccc tccgcaaaac agcctctgaa
     2341 cccaacttga aagtgcgttc aaggctaaaa cagaaggtgg ctgagcggag aagcagtccc
     2401 ctcctgcgtc gcaaggatgg gactgttatt agcaccttta agaagagagc tgttgagatc
     2461 acaggtgccg ggcctggggc gtcgtccgtg tgtaacagcg cacccggctc cggccccagc
     2521 tctcccaaca gctcccacag caccatcgct gagaatggct ttactggctc agtccccaac
     2581 atccccactg agatgctccc tcagcaccga gccctccctc tggacagctc ccccaaccag
     2641 ttcagcctct acacgtctcc ttctctgccc aacatctccc tagggctgca ggccacggtc
     2701 actgtcacca actcacacct cactgcctcc ccgaagctgt cgacacagca ggaggccgag
     2761 aggcaggccc tccagtccct gcggcagggt ggcacgctga ccggcaagtt catgagcaca
     2821 tcctctattc ctggctgcct gctgggcgtg gcactggagg gcgacgggag cccccacggg
     2881 catgcctccc tgctgcagca tgtgctgttg ctggagcagg cccggcagca gagcaccctc
     2941 attgctgtgc cactccacgg gcagtcccca ctagtgacgg gtgaacgtgt ggccaccagc
     3001 atgcggacgg taggcaagct cccgcggcat cggcccctga gccgcactca gtcctcaccg
     3061 ctgccgcaga gtccccaggc cctgcagcag ctggtcatgc aacaacagca ccagcagttc
     3121 ctggagaagc agaagcagca gcagctacag ctgggcaaga tcctcaccaa gacaggggag
     3181 ctgcccaggc agcccaccac ccaccctgag gagacagagg aggagctgac ggagcagcag
     3241 gaggtcttgc tgggggaggg agccctgacc atgccccggg agggctccac agagagtgag
     3301 agcacacagg aagacctgga ggaggaggac gaggaagagg atggggagga ggaggaggat
     3361 tgcatccagg ttaaggacga ggagggcgag agtggtgctg aggaggggcc cgacttggag
     3421 gagcctggtg ctggatacaa aaaactgttc tcagatgccc agccgctgca gcctttgcag
     3481 gtgtaccagg cgcccctcag cctggccact gtgccccacc aggccctggg ccgtacccag
     3541 tcctcccctg ctgcccctgg gggcatgaag agccccccag accagcccgt caagcacctc
     3601 ttcaccacag gtgtggtcta cgacacgttc atgctaaagc accagtgcat gtgcgggaac
     3661 acacacgtgc accctgagca tgctggccgg atccagagca tctggtcccg gctgcaggag
     3721 acaggcctgc ttagcaagtg cgagcggatc cgaggtcgca aagccacgct agatgagatc
     3781 cagacagtgc actctgaata ccacaccctg ctctatggga ccagtcccct caaccggcag
     3841 aagctagaca gcaagaagtt gctcggcccc atcagccaga agatgtatgc tgtgctgcct
     3901 tgtgggggca tcggggtgga cagtgacacc gtgtggaatg agatgcactc ctccagtgct
     3961 gtgcgcatgg cagtgggctg cctgctggag ctggccttca aggtggctgc aggagagctc
     4021 aagaatggat ttgccatcat ccggccccca ggacaccacg ccgaggaatc cacagccatg
     4081 ggattctgct tcttcaactc tgtagccatc accgcaaaac tcctacagca gaagttgaac
     4141 gtgggcaagg tcctcatcgt ggactgggac attcaccatg gcaatggcac ccagcaggcg
     4201 ttctacaatg acccctctgt gctctacatc tctctgcatc gctatgacaa cgggaacttc
     4261 tttccaggct ctggggctcc tgaagaggtt ggtggaggac caggcgtggg gtacaatgtg
     4321 aacgtggcat ggacaggagg tgtggacccc cccattggag acgtggagta ccttacagcc
     4381 ttcaggacag tggtgatgcc cattgcccac gagttctcac ctgatgtggt cctagtctcc
     4441 gccgggtttg atgctgttga aggacatctg tctcctctgg gtggctactc tgtcaccgcc
     4501 agatgttttg gccacttgac caggcagctg atgaccctgg cagggggccg ggtggtgctg
     4561 gccctggagg gaggcgctga cttgaccgcc atctgtgatg cctctgaggc ttgtgtctcg
     4621 gctctgctca gtgtagagct gcagcccttg gatgaggcag tcttgcagca aaagcccaac
     4681 atcaacgcag tggccacgct agagaaagtc atcgagatcc agagcaaaca ctggagctgt
     4741 gtgcagaagt tcgccgctgg tctgggccgg tccctgcgag aggcccaagc aggtgagacc
     4801 gaggaggccg agactgtgag cgccatggcc ttgctgtcgg tgggggccga gcaggcccag
     4861 gctgcggcag cccgggaaca cagccccagg ccggcagagg agcccatgga gcaggagcct
     4921 gccctgtgaa cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     4981 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     5041 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctgtgtg aaattgttat
     5101 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5161 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5221 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5281 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5341 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5401 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5461 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5521 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5581 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5641 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5701 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5761 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5821 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5881 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5941 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6001 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6061 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6121 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6181 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6241 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6301 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6361 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6421 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6481 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6541 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6601 tcccaacgat caaggcgagt tacatgatcc cctatgttgt gcaaaaaagc ggttagctcc
     6661 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6721 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6781 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6841 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6901 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6961 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7021 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7081 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7141 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7201 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     7261 gcactct
//
