LOCUS       sequence-225172-        6486 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 6486)
  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 6486)
  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..6486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        145..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..728
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     679..699
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     773..792
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        773..791
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      845..854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1388..1459
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1463..1483
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1484..1508
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2339..2363)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2404..2421)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2410..2634
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2680..3108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2767..2786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2977..2998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3117..3137)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3122..3451
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3302..3437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3364..3383
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3499..3546
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3565..3939
                     /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    4069..4190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4106..4125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4160..4179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4239..4255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4239..4255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4252..4274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4263..4279
                     /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(4287..4293,4294..4311,4312..4317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4332..4353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4470..4487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4641..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4721..4740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5400..6191,6192..6260))
                     /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     6023..6042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6261..6365)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6440..6459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
       61 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
      121 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      661 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      721 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
      781 ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttggta ccgagctcgg
      841 atccgccacc atggccaccc aggacgagtt cacccagctg agccagtcta tcgccgagtt
      901 ccacacctac cagctgggca acggccggtg cagcagcctg ctggcccaga gaatccacgc
      961 ccctcccgag acagtgtggt ccgtcgtgcg gagattcgac cggccccaga tctacaagca
     1021 cttcatcaag agctgcaacg tgtccgagga cttcgagatg agagtgggct gcacccggga
     1081 cgtgaacgtg atcagcggcc tgcccgccaa caccagcaga gagagactgg acctgctgga
     1141 cgacgacaga agagtgaccg gcttcagcat caccggcgga gagcaccggc tgcggaacta
     1201 caagagcgtg accaccgtgc acagattcga gaaagaggaa gaagaggaac ggatctggac
     1261 cgtggtgctg gaaagctacg tggtggacgt gcccgagggc aacagcgaag aggacacccg
     1321 gctgttcgcc gacaccgtga tccggctgaa cctgcagaag ctggcctcca tcaccgaggc
     1381 catgaacgac tacaaggacg acgacgacaa ggactataag gacgatgatg ataaggatta
     1441 caaagatgac gacgataagc cccccaagaa gaagcggaaa gtcacaagtt tgtacaaaaa
     1501 agcaggctca atgaactctc ccaacgagtc ggatgggatg tcaggtcggg aaccatcctt
     1561 ggaaatcctg ccgcggactt ctctgcacag catccctgtg acagtggagg tgaagccggt
     1621 gctgccaaga gccatgccca gttccatggg gggtgggggt ggaggcagcc ccagccctgt
     1681 ggagctacgg ggggctctgg tgggctctgt ggaccccaca ctgcgggagc agcaactgca
     1741 gcaggagctc ctggcgctca agcagcagca gcagctgcag aagcagctcc tgttcgctga
     1801 gttccagaaa cagcatgacc acctgacaag gcagcatgag gtccagctgc agaagcacct
     1861 caagcagcag caggagatgc tggcagccaa gcagcagcag gagatgctgg cagccaagcg
     1921 gcagcaggag ctggagcagc agcggcagcg ggagcagcag cggcaggaag agctggagaa
     1981 gcagcggctg gagcagcagc tgctcatcct gcggaacaag gagaagagca aagagagtgc
     2041 cattgccagc actgaggtaa agctgaggct ccaggaattc ctcttgtcga agtcaaagga
     2101 gcccacacca ggcggcctca accattccct cccacagcac cccaaatgct ggggagccca
     2161 ccatgcttct ttggaccaga gttcccctcc ccagagcggc ccccctggga cgcctccctc
     2221 ctacaaactg cctttgcctg ggccctacga cagtcgagac gacttccccc tccgcaaaac
     2281 agcctctgaa cccaacttga aagtgcgttc aaggctaaaa cagaaggtgg ctgagtagga
     2341 ccagctttct tgtacaaagt ggtcatccgc ggtctagagg gcccgtttaa acccgctgat
     2401 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2461 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2521 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2581 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2641 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2701 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2761 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2821 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2881 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2941 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3001 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3061 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3121 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3361 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3421 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3481 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3541 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     3601 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     3661 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     3721 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     3781 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     3841 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     3901 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     3961 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4021 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4081 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4141 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4201 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4261 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4321 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4381 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4441 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4501 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5041 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ctatgttgtg caaaaaagcg
     5821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6361 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     6421 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6481 tccctg
//
