LOCUS       sequence-438361-        8251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ScChd1-aa118-1274 chromatin remodeler with R126A and R130A 
            mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8251)
  AUTHORS   Nodelman IM, Das S, Faustino AM, Fried SD, Bowman GD, Armache JP
  TITLE     Nucleosome recognition and DNA distortion by the Chd1 remodeler in a
            nucleotide-free state.
  JOURNAL   Nat Struct Mol Biol. 2022 Feb;29(2):121-129. doi: 
            10.1038/s41594-021-00719-x. Epub 2022 Feb 16.
  PUBMED    35173352
REFERENCE   2  (bases 1 to 8251)
  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"; doi: 
            "10.1038/s41594-021-00719-x"; journalName: "Nat Struct Mol Biol"; 
            date: "2022-02"; volume: "29"; issue: "2"; pages: "121-129"
COMMENT     Sequence Label: pDEST17-ScChd1[R126A/R130A]
FEATURES             Location/Qualifiers
     source          1..8251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             191..214
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    complement(3720..3744)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3797..3815)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3811..3858
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4224..4229,4230..4246,4247..4252))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4279..4297)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4365..4469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4470..4538,4539..5330)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4688..4707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5501..6089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5990..6009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6243..6260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6275..6414
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6340..6359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6500..6522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6516..6707)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     8169..8188
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggctccgcgg ccgccccctt
      181 caccggatcc ctggaagttc tgttccaggg gccccaatct acagtgaaaa taccaacagc
      241 gttttctaat gctcaaaaca aaacggtaaa ttataacatc gattatagcg atgatgacct
      301 cttagaatct gaagatgact atggttccga ggaggcttta tcagaggaaa atgtgcatga
      361 agcatctgcc aatcctcaac cagaggactt ccatggtatt gatattgtta tcaatcacag
      421 actaaagaca tctttggaag aaggaaaggt cttggaaaag actgttcctg acctcaataa
      481 ctgcaaggaa aattatgaat tcttaatcaa atggacagat gaatctcatc tccacaatac
      541 ctgggaaact tatgaatcca ttggccaagt ccgtggattg aagaggttgg acaactattg
      601 taagcaattc atcatcgaag atcaacaagt aagattagat ccatacgtta ccgccgaaga
      661 tatagagatc atggacatgg aacgtgaacg taggcttgat gaatttgagg agttccacgt
      721 gccagaaagg attattgata gccaacgtgc ttcgttggaa gacggcacct cacaattaca
      781 gtatttagtt aaatggcgtc gtctaaatta cgacgaggct acttgggaga atgccacaga
      841 tatcgtgaaa ttggcacctg aacaagtgaa acattttcaa aacagagaaa actctaagat
      901 cctcccacaa tattccagta attatactag tcagagaccg cgttttgaga agttaagcgt
      961 gcaacctccg ttcattaaag gtggggaatt aagagatttt caactaactg gtattaattg
     1021 gatggcattt ttgtggtcca aaggtgataa tggtatactg gcagatgaga tgggcctggg
     1081 aaaaacggtc cagactgtcg cctttatcag ttggctgata tttgctcgta gacaaaacgg
     1141 acctcacatc attgtcgttc ctttatcgac aatgcctgcc tggttggata cttttgagaa
     1201 atgggcgcct gatttgaatt gtatatgcta tatgggcaac caaaaatcaa gagataccat
     1261 tcgagaatat gaattttaca ccaatccaag ggcaaaaggg aaaaaaacaa tgaaatttaa
     1321 tgttttatta acaacatacg agtacatctt aaaggatcgt gctgaattag gaagtataaa
     1381 atggcaattt atggccgttg acgaagctca tagactaaaa aatgctgaat catcccttta
     1441 tgaatcatta aacagtttca aggtcgccaa ccgtatgtta atcacaggca cacctcttca
     1501 gaataatatt aaagagttag ctgcgttggt taatttccta atgcccggaa ggtttacgat
     1561 tgatcaggag attgattttg aaaaccaaga tgaagagcaa gaagaatata ttcatgattt
     1621 acaccgaaga atacagcctt ttattcttcg tcggttgaag aaagacgtag aaaaatcact
     1681 tccatcaaag acagagcgta ttttaagagt tgaattgtcc gacgtacaga ctgagtacta
     1741 taaaaatatt ctgactaaaa actactctgc tttaactgct ggagctaaag ggggtcattt
     1801 ctctttactg aatattatga acgagttgaa aaaggcatcg aaccatccat atctcttcga
     1861 taatgctgaa gagcgcgtct tacagaaatt tggggatggt aaaatgactc gagaaaacgt
     1921 actaagaggt ttgatcatgt cttcgggtaa gatggttctt ttagaccaat tattgaccag
     1981 attgaagaaa gatgggcacc gcgtgttgat tttttcacaa atggtcagaa tgcttgacat
     2041 tttaggcgac tatttatcca ttaaaggtat taacttccaa aggttagatg gtacagtacc
     2101 atctgctcaa agaaggatat ctattgatca tttcaactct ccggattcaa acgacttcgt
     2161 atttttactt tctactcgtg ctggtggttt gggtatcaac ttaatgaccg ccgataccgt
     2221 tgtgattttt gattccgatt ggaatccgca agccgattta caagcaatgg ctagagctca
     2281 tcgtattggc caaaaaaatc acgttatggt gtataggttg gtttcaaaag acacagtaga
     2341 ggaagaggta ttagaaagag cacggaagaa aatgattctg gaatatgcta ttatttctct
     2401 tggagtgaca gatggtaaca aatacactaa gaagaatgaa ccaaatgccg gtgaattatc
     2461 ggcaatcctg aagttcggtg caggcaatat gttcactgcc acagacaacc agaaaaaatt
     2521 ggaggatctt aacttggatg atgttttgaa tcatgcagag gatcacgtta ctactccaga
     2581 cttaggagag tcgcatcttg gcggtgaaga gtttttgaag caattcgaag tgactgatta
     2641 taaagctgat atagattggg atgatatcat tccagaagaa gaactaaaaa aactccaaga
     2701 tgaagagcag aaacgcaagg atgaagaata tgttaaagaa caacttgaaa tgatgaatag
     2761 aagggataac gccctaaaaa aaataaaaaa cagtgttaat ggtgatggga ccgccgcaaa
     2821 ctcagattct gacgacgata gtacttccag atcttctaga agaagagcaa gagctaatga
     2881 catggactct attggtgaat cggaggtgag agctttgtat aaagctattt taaaatttgg
     2941 caacctaaag gagattttgg acgagttgat tgcagatgga accctgccgg tcaaatcatt
     3001 tgaaaaatac ggtgaaactt acgatgaaat gatggaagca gctaaagact gtgtacacga
     3061 ggaagagaaa aatagaaaag aaatcttgga gaaacttgag aagcatgcta ccgcctatag
     3121 agcaaagcta aaaagcggtg aaataaaagc agagaaccaa ccaaaggata atccgttgac
     3181 tagattatct ttaaaaaaaa gagaaaagaa agccgtcctt ttcaacttta aaggtgttaa
     3241 atctttaaac gccgaatctt tactaagtag agtggaggat ttaaagtact tgaagaactt
     3301 gataaattca aactacaaag atgatccatt aaagtttagc ctaggcaaca acacacctaa
     3361 gcctgtacag aattggtcat ctaattggac gaaagaggaa gatgagaagc tattgattgg
     3421 tgttttcaaa tatggatatg gttcctggac acaaataaga gacgatccat ttctaggcat
     3481 tactgataaa atattcttga atgaagttca caatccggta gcaaaaaagt cggcaagctc
     3541 ctctgataca acaccaacac cctcaaagaa aggaaaggga ataacgggtt cttcaaaaaa
     3601 agtacctggt gccattcact tgggcagaag agttgattat ctattatcct tcttaagagg
     3661 aggcctaaat acaaagagtc ccagttgaaa gcttaataat aataagggtg ggcgcgccga
     3721 cccagctttc ttgtacaaag tggttgattc gaggctgcta acaaagcccg aaaggaagct
     3781 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     3841 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatccaca ggacgggtgt
     3901 ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc gaagcgagca ggactgggcg
     3961 gcggccaaag cggtcggaca gtgctccgag aacgggtgcg catagaaatt gcatcaacgc
     4021 atatagcgct agcagcacgc catagtgact ggcgatgctg tcggaatgga cgatatcccg
     4081 caagaggccc ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt
     4141 gccgaggatg acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca
     4201 atttaactgt gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga
     4261 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4321 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4681 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4981 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     5041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5641 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6241 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     6301 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     6361 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     6421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6541 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     6601 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     6661 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     6721 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     6781 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     6841 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     6901 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     6961 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     7021 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     7081 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     7141 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     7201 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     7261 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     7321 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     7381 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     7441 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta
     7501 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     7561 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     7621 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     7681 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     7741 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     7801 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     7861 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     7921 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga tcgacgctct
     7981 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     8041 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     8101 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     8161 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     8221 ccggccacga tgcgtccggc gtagaggatc g
//
