LOCUS       sequence-225208-        8935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PYL-HDAC5 catalytic domain swap to HDAC3 catalytic domain 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8935)
  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 8935)
  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(EXPR-DS-HDAC5_HDAC3)
FEATURES             Location/Qualifiers
     source          1..8935
                     /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"
     primer_bind     823..842
                     /label=T7
                     /note="T7 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(4838..4862)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4903..4920)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4909..5133
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5179..5607
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5266..5285)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5476..5497
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5616..5636)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5621..5950
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5801..5936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5863..5882
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5998..6045
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6064..6438
                     /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    6568..6689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6605..6624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6659..6678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6738..6754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6738..6754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6751..6773)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6762..6778
                     /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(6786..6792,6793..6810,6811..6816))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6831..6852
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6969..6986)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7140..7728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7220..7239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7899..8690,8691..8759))
                     /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     8522..8541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8760..8864)
                     /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 agcaggcttc
     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 ttcaccacaa agaccgtggc ctatttctac gaccccgacg tgggcaactt ccactacgga
     3661 gctggacacc ctatgaagcc ccatcgcctg gcattgaccc atagcctggt cctgcattac
     3721 ggtctctata agaagatgat cgtcttcaag ccataccagg cctcccagca tgacatgtgc
     3781 cgcttccact ccgaggacta cattgacttc ctgcagagag tcagccccac caatatgcaa
     3841 ggcttcacca agagtcttaa tgccttcaac gtaggcgatg actgcccagt gtttcccggg
     3901 ctctttgagt tctgctcgcg ttacacaggc gcatctctgc aaggagcaac ccagctgaac
     3961 aacaagatct gtgatattgc cattaactgg gctggtggtc tgcaccatgc caagaagttt
     4021 gaggcctctg gcttctgcta tgtcaacgac attgtgattg gcatcctgga gctgctcaag
     4081 taccaccctc gggtgctcta cattgacatt gacatccacc atggtgacgg ggttcaagaa
     4141 gctttctacc tcactgaccg ggtcatgacg gtgtccttcc acaaatacgg aaattacttc
     4201 ttccctggca caggtgacat gtatgaagtc ggggcagaga gtggccgcta ctactgtctg
     4261 aacgtgcccc tgcgggatgg cattgatgac cagagttaca agcacctttt ccagccggtt
     4321 atcaaccagg tagtggactt ctaccaaccc acgtgcattg tgctccagtg tggagctgac
     4381 tctctgggct gtgatcgatt gggctgcttt aacctcagca tccgagggca tggggaatgc
     4441 gttgaatatg tcaagagctt caatatccct ctactcgtgc tgggtggtgg tggttatact
     4501 gtccgaaatg ttgcccgctg ctggacatat gagacatcgc tgctggtaga acccttggat
     4561 gaggcagtct tgcagcaaaa gcccaacatc aacgcagtgg ccacgctaga gaaagtcatc
     4621 gagatccaga gcaaacactg gagctgtgtg cagaagttcg ccgctggtct gggccggtcc
     4681 ctgcgagagg cccaagcagg tgagaccgag gaggccgaga ctgtgagcgc catggccttg
     4741 ctgtcggtgg gggccgagca ggcccaggct gcggcagccc gggaacacag ccccaggccg
     4801 gcagaggagc ccatggagca ggagcctgcc ctgtgagacc cagctttctt gtacaaagtg
     4861 gtcatccgcg gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     4921 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4981 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5041 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5101 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     5161 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5221 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5281 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5341 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5401 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5461 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5521 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5581 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     5641 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5701 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5761 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5821 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5881 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5941 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     6001 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     6061 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     6121 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     6181 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     6241 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     6301 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     6361 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     6421 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
     6481 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6541 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6601 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6661 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6721 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6781 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6841 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6901 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6961 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7021 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7081 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7141 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7201 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7261 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7321 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7381 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7441 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7501 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7561 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7621 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7681 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7741 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7801 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7861 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7921 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7981 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8041 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8101 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8161 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8221 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8281 aggcgagtta catgatcccc tatgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8341 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8401 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8461 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8521 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8581 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8641 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8701 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8761 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8821 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8881 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actct
//
