LOCUS       sequence-215892-        8936 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 8936)
  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 8936)
  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..8936
                     /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(4878..4902)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4943..4960)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4949..5173
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5219..5647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5306..5325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5516..5537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5656..5676)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5661..5991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5842..5977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5904..5923
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6039..6086
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6105..6479
                     /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    6609..6730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6646..6665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6700..6719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6779..6795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6779..6795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6792..6814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6803..6819
                     /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(6827..6833,6834..6851,6852..6857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6872..6893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7010..7027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7181..7769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7261..7280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7940..8731,8732..8800))
                     /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     8563..8582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8801..8905)
                     /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 agcaggcttc 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 ttcaccacaa agaccgtggc
     3661 ctatttctac gaccccgacg tgggcaactt ccactacgga gctggacacc ctatgaagcc
     3721 ccatcgcctg gcattgaccc atagcctggt cctgcattac ggtctctata agaagatgat
     3781 cgtcttcaag ccataccagg cctcccagca tgacatgtgc cgcttccact ccgaggacta
     3841 cattgacttc ctgcagagag tcagccccac caatatgcaa ggcttcacca agagtcttaa
     3901 tgccttcaac gtaggcgatg actgcccagt gtttcccggg ctctttgagt tctgctcgcg
     3961 ttacacaggc gcatctctgc aaggagcaac ccagctgaac aacaagatct gtgatattgc
     4021 cattaactgg gctggtggtc tgcaccatgc caagaagttt gaggcctctg gcttctgcta
     4081 tgtcaacgac attgtgattg gcatcctgga gctgctcaag taccaccctc gggtgctcta
     4141 cattgacatt gacatccacc atggtgacgg ggttcaagaa gctttctacc tcactgaccg
     4201 ggtcatgacg gtgtccttcc acaaatacgg aaattacttc ttccctggca caggtgacat
     4261 gtatgaagtc ggggcagaga gtggccgcta ctactgtctg aacgtgcccc tgcgggatgg
     4321 cattgatgac cagagttaca agcacctttt ccagccggtt atcaaccagg tagtggactt
     4381 ctaccaaccc acgtgcattg tgctccagtg tggagctgac tctctgggct gtgatcgatt
     4441 gggctgcttt aacctcagca tccgagggca tggggaatgc gttgaatatg tcaagagctt
     4501 caatatccct ctactcgtgc tgggtggtgg tggttatact gtccgaaatg ttgcccgctg
     4561 ctggacatat gagacatcgc tgctggtaga acccttggat gaggcagtct tgcagcaaaa
     4621 gcccaacatc aacgcagtgg ccacgctaga gaaagtcatc gagatccaga gcaaacactg
     4681 gagctgtgtg cagaagttcg ccgctggtct gggccggtcc ctgcgagagg cccaagcagg
     4741 tgagaccgag gaggccgaga ctgtgagcgc catggccttg ctgtcggtgg gggccgagca
     4801 ggcccaggct gcggcagccc gggaacacag ccccaggccg gcagaggagc ccatggagca
     4861 ggagcctgcc ctgtgagacc cagctttctt gtacaaagtg gtcatccgcg gtctagaggg
     4921 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4981 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     5041 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     5101 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     5161 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     5221 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5281 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5341 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     5401 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5461 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5521 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5581 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5641 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6001 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     6061 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
     6121 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     6181 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     6241 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     6301 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     6361 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     6421 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     6481 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6541 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6601 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6661 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6721 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6781 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6841 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6901 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     7381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7621 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
