LOCUS       sequence-225168-        7711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PYL-HDAC5 N terminal domain expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  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 7711)
  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(PYL-Nterm_dom_HDAC5)
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        142..521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        522..725
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     676..696
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     770..789
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        770..788
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      842..851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1385..1456
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1460..1480
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1481..1505
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3561..3585)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3626..3643)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3632..3856
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3902..4330
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3989..4008)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4199..4220
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4339..4359)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4344..4673
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4524..4659
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4586..4605
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4721..4768
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4787..5161
                     /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    5291..5412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5328..5347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5382..5401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5461..5477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5461..5477)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5474..5496)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5485..5501
                     /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(5509..5515,5516..5533,5534..5539))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5554..5575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5692..5709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5863..6451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5943..5962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6622..7413,7414..7482))
                     /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     7245..7264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7483..7587)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7662..7681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
       61 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      121 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      181 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      241 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      301 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      361 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      421 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      481 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      541 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      601 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      661 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      721 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      781 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      841 cgccaccatg gccacccagg acgagttcac ccagctgagc cagtctatcg ccgagttcca
      901 cacctaccag ctgggcaacg gccggtgcag cagcctgctg gcccagagaa tccacgcccc
      961 tcccgagaca gtgtggtccg tcgtgcggag attcgaccgg ccccagatct acaagcactt
     1021 catcaagagc tgcaacgtgt ccgaggactt cgagatgaga gtgggctgca cccgggacgt
     1081 gaacgtgatc agcggcctgc ccgccaacac cagcagagag agactggacc tgctggacga
     1141 cgacagaaga gtgaccggct tcagcatcac cggcggagag caccggctgc ggaactacaa
     1201 gagcgtgacc accgtgcaca gattcgagaa agaggaagaa gaggaacgga tctggaccgt
     1261 ggtgctggaa agctacgtgg tggacgtgcc cgagggcaac agcgaagagg acacccggct
     1321 gttcgccgac accgtgatcc ggctgaacct gcagaagctg gcctccatca ccgaggccat
     1381 gaacgactac aaggacgacg acgacaagga ctataaggac gatgatgata aggattacaa
     1441 agatgacgac gataagcccc ccaagaagaa gcggaaagtc acaagtttgt acaaaaaagc
     1501 aggctcaatg aactctccca acgagtcgga tgggatgtca ggtcgggaac catccttgga
     1561 aatcctgccg cggacttctc tgcacagcat ccctgtgaca gtggaggtga agccggtgct
     1621 gccaagagcc atgcccagtt ccatgggggg tgggggtgga ggcagcccca gccctgtgga
     1681 gctacggggg gctctggtgg gctctgtgga ccccacactg cgggagcagc aactgcagca
     1741 ggagctcctg gcgctcaagc agcagcagca gctgcagaag cagctcctgt tcgctgagtt
     1801 ccagaaacag catgaccacc tgacaaggca gcatgaggtc cagctgcaga agcacctcaa
     1861 gcagcagcag gagatgctgg cagccaagca gcagcaggag atgctggcag ccaagcggca
     1921 gcaggagctg gagcagcagc ggcagcggga gcagcagcgg caggaagagc tggagaagca
     1981 gcggctggag cagcagctgc tcatcctgcg gaacaaggag aagagcaaag agagtgccat
     2041 tgccagcact gaggtaaagc tgaggctcca ggaattcctc ttgtcgaagt caaaggagcc
     2101 cacaccaggc ggcctcaacc attccctccc acagcacccc aaatgctggg gagcccacca
     2161 tgcttctttg gaccagagtt cccctcccca gagcggcccc cctgggacgc ctccctccta
     2221 caaactgcct ttgcctgggc cctacgacag tcgagacgac ttccccctcc gcaaaacagc
     2281 ctctgaaccc aacttgaaag tgcgttcaag gctaaaacag aaggtggctg agcggagaag
     2341 cagtcccctc ctgcgtcgca aggatgggac tgttattagc acctttaaga agagagctgt
     2401 tgagatcaca ggtgccgggc ctggggcgtc gtccgtgtgt aacagcgcac ccggctccgg
     2461 ccccagctct cccaacagct cccacagcac catcgctgag aatggcttta ctggctcagt
     2521 ccccaacatc cccactgaga tgctccctca gcaccgagcc ctccctctgg acagctcccc
     2581 caaccagttc agcctctaca cgtctccttc tctgcccaac atctccctag ggctgcaggc
     2641 cacggtcact gtcaccaact cacacctcac tgcctccccg aagctgtcga cacagcagga
     2701 ggccgagagg caggccctcc agtccctgcg gcagggtggc acgctgaccg gcaagttcat
     2761 gagcacatcc tctattcctg gctgcctgct gggcgtggca ctggagggcg acgggagccc
     2821 ccacgggcat gcctccctgc tgcagcatgt gctgttgctg gagcaggccc ggcagcagag
     2881 caccctcatt gctgtgccac tccacgggca gtccccacta gtgacgggtg aacgtgtggc
     2941 caccagcatg cggacggtag gcaagctccc gcggcatcgg cccctgagcc gcactcagtc
     3001 ctcaccgctg ccgcagagtc cccaggccct gcagcagctg gtcatgcaac aacagcacca
     3061 gcagttcctg gagaagcaga agcagcagca gctacagctg ggcaagatcc tcaccaagac
     3121 aggggagctg cccaggcagc ccaccaccca ccctgaggag acagaggagg agctgacgga
     3181 gcagcaggag gtcttgctgg gggagggagc cctgaccatg ccccgggagg gctccacaga
     3241 gagtgagagc acacaggaag acctggagga ggaggacgag gaagaggatg gggaggagga
     3301 ggaggattgc atccaggtta aggacgagga gggcgagagt ggtgctgagg aggggcccga
     3361 cttggaggag cctggtgctg gatacaaaaa actgttctca gatgcccagc cgctgcagcc
     3421 tttgcaggtg taccaggcgc ccctcagcct ggccactgtg ccccaccagg ccctgggccg
     3481 tacccagtcc tcccctgctg cccctggggg catgaagagc cccccagacc agcccgtcaa
     3541 gcacctcttc accacatagg acccagcttt cttgtacaaa gtggtcatcc gcggtctaga
     3601 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3661 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3721 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3781 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3841 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3901 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3961 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4021 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4081 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4141 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4201 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4261 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4321 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4381 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4441 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4501 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4561 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4621 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4681 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4741 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     4801 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     4861 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     4921 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     4981 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     5041 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     5101 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     5161 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5221 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5281 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5341 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5401 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5461 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5521 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5581 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5641 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5701 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5821 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5881 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5941 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6001 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6061 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6121 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6181 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6241 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6301 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6361 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6421 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6481 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6541 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6601 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6661 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6721 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6781 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6841 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6901 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6961 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7021 ccctatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7081 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7141 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7201 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7261 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7321 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7381 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7441 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7501 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7561 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7621 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7681 gcatagttaa gccagtatct gctccctgct t
//
