LOCUS       sequence-225202-        6697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For PYL-HDAC5 catalytic domain expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  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 6697)
  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-cat_dom_HDAC5)
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     695..715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     789..808
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        789..807
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1404..1475
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1479..1499
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1500..1524
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2566..2590)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2631..2648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2637..2861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2907..3335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2994..3013)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3204..3225
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3344..3364)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3349..3678
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3529..3664
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3591..3610
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3726..3773
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3792..4166
                     /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    4296..4417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4333..4352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4387..4406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4466..4482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4466..4482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4479..4501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4490..4506
                     /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(4514..4520,4521..4538,4539..4544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4559..4580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4697..4714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4868..5456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4948..4967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5627..6418,6419..6487))
                     /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     6250..6269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6488..6592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6667..6686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
       61 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
      121 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      781 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
      841 ttggtaccga gctcggatcc gccaccatgg ccacccagga cgagttcacc cagctgagcc
      901 agtctatcgc cgagttccac acctaccagc tgggcaacgg ccggtgcagc agcctgctgg
      961 cccagagaat ccacgcccct cccgagacag tgtggtccgt cgtgcggaga ttcgaccggc
     1021 cccagatcta caagcacttc atcaagagct gcaacgtgtc cgaggacttc gagatgagag
     1081 tgggctgcac ccgggacgtg aacgtgatca gcggcctgcc cgccaacacc agcagagaga
     1141 gactggacct gctggacgac gacagaagag tgaccggctt cagcatcacc ggcggagagc
     1201 accggctgcg gaactacaag agcgtgacca ccgtgcacag attcgagaaa gaggaagaag
     1261 aggaacggat ctggaccgtg gtgctggaaa gctacgtggt ggacgtgccc gagggcaaca
     1321 gcgaagagga cacccggctg ttcgccgaca ccgtgatccg gctgaacctg cagaagctgg
     1381 cctccatcac cgaggccatg aacgactaca aggacgacga cgacaaggac tataaggacg
     1441 atgatgataa ggattacaaa gatgacgacg ataagccccc caagaagaag cggaaagtca
     1501 caagtttgta caaaaaagca ggcttcggtg tggtctacga cacgttcatg ctaaagcacc
     1561 agtgcatgtg cgggaacaca cacgtgcacc ctgagcatgc tggccggatc cagagcatct
     1621 ggtcccggct gcaggagaca ggcctgctta gcaagtgcga gcggatccga ggtcgcaaag
     1681 ccacgctaga tgagatccag acagtgcact ctgaatacca caccctgctc tatgggacca
     1741 gtcccctcaa ccggcagaag ctagacagca agaagttgct cggccccatc agccagaaga
     1801 tgtatgctgt gctgccttgt gggggcatcg gggtggacag tgacaccgtg tggaatgaga
     1861 tgcactcctc cagtgctgtg cgcatggcag tgggctgcct gctggagctg gccttcaagg
     1921 tggctgcagg agagctcaag aatggatttg ccatcatccg gcccccagga caccacgccg
     1981 aggaatccac agccatggga ttctgcttct tcaactctgt agccatcacc gcaaaactcc
     2041 tacagcagaa gttgaacgtg ggcaaggtcc tcatcgtgga ctgggacatt caccatggca
     2101 atggcaccca gcaggcgttc tacaatgacc cctctgtgct ctacatctct ctgcatcgct
     2161 atgacaacgg gaacttcttt ccaggctctg gggctcctga agaggttggt ggaggaccag
     2221 gcgtggggta caatgtgaac gtggcatgga caggaggtgt ggaccccccc attggagacg
     2281 tggagtacct tacagccttc aggacagtgg tgatgcccat tgcccacgag ttctcacctg
     2341 atgtggtcct agtctccgcc gggtttgatg ctgttgaagg acatctgtct cctctgggtg
     2401 gctactctgt caccgccaga tgttttggcc acttgaccag gcagctgatg accctggcag
     2461 ggggccgggt ggtgctggcc ctggagggag gccatgactt gaccgccatc tgtgatgcct
     2521 ctgaggcttg tgtctcggct ctgctcagtg tagagctgca gtaggaccca gctttcttgt
     2581 acaaagtggt catccgcggt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     2641 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2701 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2761 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2821 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2881 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2941 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3001 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3061 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3121 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3181 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3241 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3301 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3361 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3601 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3661 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     3721 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3781 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     3841 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     3901 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     3961 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     4021 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     4081 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     4141 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     4201 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4261 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4441 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4621 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4681 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4741 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4801 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4861 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4921 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4981 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5041 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5101 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5161 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5221 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5281 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5341 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5401 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5461 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5521 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5581 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5761 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6001 aacgatcaag gcgagttaca tgatccccta tgttgtgcaa aaaagcggtt agctccttcg
     6061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6601 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6661 tctcagtaca atctgctctg atgccgcata gttaagc
//
