LOCUS       sequence-342746-        8415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ALC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8415)
  AUTHORS   Ooi SK, Sato S, Tomomori-Sato C, Zhang Y, Wen Z, Banks CAS, Washburn
            MP, Unruh JR, Florens L, Conaway RC, Conaway JW
  TITLE     Multiple roles for PARP1 in ALC1-dependent nucleosome remodeling.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Sep 7;118(36). pii: 2107277118. doi: 
            10.1073/pnas.2107277118.
  PUBMED    34465625
REFERENCE   2  (bases 1 to 8415)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-09-7"; volume: "118(36). pii: 
            2107277118. doi"; pages: " 10.1073/pnas.2107277118"
COMMENT     Sequence Label: pET41sIII_GST-ALC1
FEATURES             Location/Qualifiers
     source          1..8415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..1083)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1045..1064
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1084..1161)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1367..1386
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1474..1493
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1474..1492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1493..1517
                     /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)."
     RBS             1532..1554
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1562..2215
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2173..2195
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2234..2257
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4970..4993
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(5056..5074)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5070..5117
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5154..5609
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5241..5260)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5451..5472
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5701..6516)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5701..6516)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6424..6443
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6508..6527
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6638..7226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7127..7146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7380..7397
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7412..7551
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7477..7496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7637..7659
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7653..7844)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
       61 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
      121 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
      181 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
      241 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
      301 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
      361 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
      421 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
      481 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
      541 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
      601 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
      661 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
      721 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
      781 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
      841 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
      901 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
      961 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     1021 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     1081 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     1141 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     1201 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     1261 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1321 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1381 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1441 agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt
     1501 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
     1561 tatgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
     1621 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
     1681 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
     1741 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
     1801 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
     1861 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
     1921 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
     1981 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
     2041 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
     2101 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
     2161 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga
     2221 tggttcaact agtctggaag ttctgttcca ggggcccctc gagatggagc gcgcgggcgc
     2281 tactagccgc gggggccagg cgcctggctt cttactgcgg cttcatactg agggcccagc
     2341 cgaggcggcg cgggtgcagg agcaggactt acggcagtgg gggctgacag ggattcacct
     2401 acgctcttac cagctggagg gagtaaactg gctcgcccag cgcttccatt gtcagaatgg
     2461 ctgtatcctg ggagatgaga tgggcctggg gaagacctgc cagactattg ctctcttcat
     2521 ttatttggca ggaagattaa atgatgaagg gccatttctg attctttgtc ccttgtctgt
     2581 tttgagcaac tggaaagaag aaatgcagag atttgctcca ggtctttcct gtgtaacata
     2641 tgcaggcgac aaggaggaaa gagcctgcct tcagcaagac ctgaaacagg agtcacgttt
     2701 tcatgtgcta ctgactacct atgagatttg cttgaaagat gcatcatttc taaaatcatt
     2761 cccttggagt gttcttgttg tggatgaagc tcacaggttg aaaaaccaaa gctccctgct
     2821 gcataagacc ttgtcagagt tctcagtagt cttcagtctc ctgttgaccg gaactcccat
     2881 ccagaacagc ctccaagagc tctactccct cctcagtttt gtggagcctg atctcttttc
     2941 caaggaagag gtgggagatt ttattcaacg ctaccaggat attgagaaag aatctgagtc
     3001 agcaagtgaa ctgcacaaac tcttgcagcc atttctgctg aggcgagtga aagctgaggt
     3061 agctacagag cttcccaaga agacagaagt agtgatatac catggcatgt cagcattgca
     3121 gaagaaatac tacaaggcca ttttgatgaa agacctagat gcatttgaaa atgagacggc
     3181 aaagaaagtt aaactacaga acattttgtc ccagcttcga aagtgtgtgg atcacccata
     3241 tttgtttgat ggtgtggagc cggagccttt tgaagttgga gaccacctga ctgaggctag
     3301 tgggaagctt cacctgctgg ataagctact agcattcctg tattctgggg gccatcgggt
     3361 tttacttttc tcccaaatga cccagatgtt ggatattctc caagactata tggattacag
     3421 aggctacagc tatgagcgtg tggatggttc tgtgagagga gaagagagac acttggccat
     3481 taagaacttt ggacagcagc ccattttcgt ttttctcctg agtactaggg caggtggagt
     3541 tggcatgaac ttaacagcag cagatactgt gatttttgtt gacagtgact ttaatcctca
     3601 gaatgacttg caagcagctg ccagggctca tcgcattggc caaaacaagt ctgttaaagt
     3661 tattcggctg attggtcgag acactgtgga agaaatagtc tataggaaag cagcctccaa
     3721 actgcagctc accaacatga tcatagaagg aggccatttt actctgggag cccaaaaacc
     3781 cgctgccgat gctgacctcc agttgagtga gatactcaaa tttggtttgg ataaactgct
     3841 ggcctctgag gggagcacca tggatgaaat agacctggag tccatcctgg gagaaacaaa
     3901 agatggccag tgggtctctg atgccttgcc tgcagcagaa ggagggagca gagatcaaga
     3961 ggaaggaaaa aatcatatgt acttatttga aggtaaagat tattctaaag agcccagtaa
     4021 ggaagacaga aaatcatttg aacaactggt aaaccttcag aaaacccttt tggagaaagc
     4081 tagtcaagag ggccgatcac tccgaaataa aggcagtgtt ctcatcccag gccttgtgga
     4141 gggatctacc aaaaggaagc gggttctgag tccagaagag ctggaggaca gacagaagaa
     4201 aagacaagaa gcagctgcca agagaaggag actcatagag gagaagaaga ggcaaaagga
     4261 agaggctgaa cataagaaaa agatggcctg gtgggaatcc aacaattacc agtccttctg
     4321 cctgccctct gaggagagcg agccagagga ccttgagaat ggggaagaga gctctgctga
     4381 gctggattac caagacccag atgctacttc cctcaagtac gttagtggtg atgtcaccca
     4441 ccctcaggct ggggccgagg atgctctcat tgtgcactgc gtagatgact gtggccactg
     4501 gggcagaggt ggtttattta cagctctgga aaagcgatcc gctgagccaa gaaaaatata
     4561 tgagctggct gggaaaatga aagacctgag tttgggaggt gtccttttat ttcctgttga
     4621 tgataaagaa tcaagaaaca aagggcaaga tttgttggcc ttgattgtgg ctcagcatcg
     4681 tgatcgttcc aatgtcctgt ctggcattaa gatggcagcc ctagaagagg gcctgaagaa
     4741 gatattttta gcagcaaaaa agaagaaagc aagtgttcat cttccacgta ttggacatgc
     4801 cacgaaaggt tttaactggt atggtactga gcgacttatt cggaaacatc tggctgcaag
     4861 aggcatccca acttacatat attattttcc tagaagcaag tctgctgtcc ttcattcaca
     4921 gtcttcatct tcctcctcaa gacagctggt gccttaagga tccgtcgagc accaccacca
     4981 ccaccaccac cactaattga ttaataccta ggctgctaaa caaagcccga aaggaagctg
     5041 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     5101 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     5161 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5221 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5281 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5341 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5401 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5461 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5521 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5581 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     5701 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     5761 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     5821 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     5881 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     5941 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     6001 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     6061 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     6121 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     6181 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     6241 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     6301 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     6361 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     6421 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     6481 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6541 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     6601 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6661 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6721 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6781 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6841 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6901 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6961 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7021 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7081 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7141 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7201 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7261 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7321 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7381 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     7441 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     7501 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     7561 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     7621 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     7681 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     7741 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     7801 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     7861 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     7921 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     7981 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     8041 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     8101 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     8161 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     8221 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     8281 aacgacagga gcacgatcat gctagtcatg ccccgcgccc accggaagga gctgactggg
     8341 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     8401 ttaattgcgt tgcgc
//
