LOCUS       sequence-215893-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PYL-HDAC5 aa 1-275 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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 6488)
  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-HDAC5-trun1)
FEATURES             Location/Qualifiers
     source          1..6488
                     /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(2430..2454)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2495..2512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2501..2725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2771..3199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2858..2877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3068..3089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3208..3228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3213..3543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3394..3529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3456..3475
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3591..3638
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3657..4031
                     /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    4161..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4198..4217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4252..4271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4331..4347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4331..4347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4344..4366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4355..4371
                     /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(4379..4385,4386..4403,4404..4409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4424..4445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4562..4579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4733..5321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4813..4832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5492..6283,6284..6352))
                     /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     6115..6134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6353..6457)
                     /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 ctgagtagga cccagctttc ttgtacaaag tggtcatccg
     2461 cggtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2521 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2581 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2641 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2701 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2761 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2821 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2881 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     2941 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3001 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3061 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3121 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     3181 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3241 caggctcccc aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3481 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3541 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     3601 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     3661 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     3721 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     3781 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     3841 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     3901 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     3961 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     4021 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4081 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4141 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4201 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4261 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4321 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4381 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4681 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4741 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4801 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4861 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4921 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5161 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5341 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5401 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5461 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5521 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5581 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5641 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5701 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5761 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5821 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5881 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5941 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6001 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6061 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6121 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6181 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6241 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6301 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6361 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6421 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6481 ctgacgtc
//
