LOCUS       sequence-343400-        8415 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Bacterial expression of inactive ALC1 (E175Q disrupts ATP binding).
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 Apr 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-E175Q
FEATURES             Location/Qualifiers
     source          1..8415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             89..742
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     700..722
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             761..784
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3497..3520
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3583..3601)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3597..3644
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3681..4136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3768..3787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3978..3999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4228..5043)
                     /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(4228..5043)
                     /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     4951..4970
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5035..5054
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5165..5753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5654..5673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5907..5924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5939..6078
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6004..6023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6164..6186
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6180..6371)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6943..8025)
                     /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     7987..8006
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8026..8103)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8309..8328
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtcccctata ctaggttatt ggaaaattaa
      121 gggccttgtg caacccactc gacttctttt ggaatatctt gaagaaaaat atgaagagca
      181 tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat tgggtttgga
      241 gtttcccaat cttccttatt atattgatgg tgatgttaaa ttaacacagt ctatggccat
      301 catacgttat atagctgaca agcacaacat gttgggtggt tgtccaaaag agcgtgcaga
      361 gatttcaatg cttgaaggag cggttttgga tattagatac ggtgtttcga gaattgcata
      421 tagtaaagac tttgaaactc tcaaagttga ttttcttagc aagctacctg aaatgctgaa
      481 aatgttcgaa gatcgtttat gtcataaaac atatttaaat ggtgatcatg taacccatcc
      541 tgacttcatg ttgtatgacg ctcttgatgt tgttttatac atggacccaa tgtgcctgga
      601 tgcgttccca aaattagttt gttttaaaaa acgtattgaa gctatcccac aaattgataa
      661 gtacttgaaa tccagcaagt atatagcatg gcctttgcag ggctggcaag ccacgtttgg
      721 tggtggcgac catcctccaa aatcggatgg ttcaactagt ctggaagttc tgttccaggg
      781 gcccctcgag atggagcgcg cgggcgctac tagccgcggg ggccaggcgc ctggcttctt
      841 actgcggctt catactgagg gcccagccga ggcggcgcgg gtgcaggagc aggacttacg
      901 gcagtggggg ctgacaggga ttcacctacg ctcttaccag ctggagggag taaactggct
      961 cgcccagcgc ttccattgtc agaatggctg tatcctggga gatgagatgg gcctggggaa
     1021 gacctgccag actattgctc tcttcattta tttggcagga agattaaatg atgaagggcc
     1081 atttctgatt ctttgtccct tgtctgtttt gagcaactgg aaagaagaaa tgcagagatt
     1141 tgctccaggt ctttcctgtg taacatatgc aggcgacaag gaggaaagag cctgccttca
     1201 gcaagacctg aaacaggagt cacgttttca tgtgctactg actacctatg agatttgctt
     1261 gaaagatgca tcatttctaa aatcattccc ttggagtgtt cttgttgtgg atcaggctca
     1321 caggttgaaa aaccaaagct ccctgctgca taagaccttg tcagagttct cagtagtctt
     1381 cagtctcctg ttgaccggaa ctcccatcca gaacagcctc caagagctct actccctcct
     1441 cagttttgtg gagcctgatc tcttttccaa ggaagaggtg ggagatttta ttcaacgcta
     1501 ccaggatatt gagaaagaat ctgagtcagc aagtgaactg cacaaactct tgcagccatt
     1561 tctgctgagg cgagtgaaag ctgaggtagc tacagagctt cccaagaaga cagaagtagt
     1621 gatataccat ggcatgtcag cattgcagaa gaaatactac aaggccattt tgatgaaaga
     1681 cctagatgca tttgaaaatg agacggcaaa gaaagttaaa ctacagaaca ttttgtccca
     1741 gcttcgaaag tgtgtggatc acccatattt gtttgatggt gtggagccgg agccttttga
     1801 agttggagac cacctgactg aggctagtgg gaagcttcac ctgctggata agctactagc
     1861 attcctgtat tctgggggcc atcgggtttt acttttctcc caaatgaccc agatgttgga
     1921 tattctccaa gactatatgg attacagagg ctacagctat gagcgtgtgg atggttctgt
     1981 gagaggagaa gagagacact tggccattaa gaactttgga cagcagccca ttttcgtttt
     2041 tctcctgagt actagggcag gtggagttgg catgaactta acagcagcag atactgtgat
     2101 ttttgttgac agtgacttta atcctcagaa tgacttgcaa gcagctgcca gggctcatcg
     2161 cattggccaa aacaagtctg ttaaagttat tcggctgatt ggtcgagaca ctgtggaaga
     2221 aatagtctat aggaaagcag cctccaaact gcagctcacc aacatgatca tagaaggagg
     2281 ccattttact ctgggagccc aaaaacccgc tgccgatgct gacctccagt tgagtgagat
     2341 actcaaattt ggtttggata aactgctggc ctctgagggg agcaccatgg atgaaataga
     2401 cctggagtcc atcctgggag aaacaaaaga tggccagtgg gtctctgatg ccttgcctgc
     2461 agcagaagga gggagcagag atcaagagga aggaaaaaat catatgtact tatttgaagg
     2521 taaagattat tctaaagagc ccagtaagga agacagaaaa tcatttgaac aactggtaaa
     2581 ccttcagaaa acccttttgg agaaagctag tcaagagggc cgatcactcc gaaataaagg
     2641 cagtgttctc atcccaggcc ttgtggaggg atctaccaaa aggaagcggg ttctgagtcc
     2701 agaagagctg gaggacagac agaagaaaag acaagaagca gctgccaaga gaaggagact
     2761 catagaggag aagaagaggc aaaaggaaga ggctgaacat aagaaaaaga tggcctggtg
     2821 ggaatccaac aattaccagt ccttctgcct gccctctgag gagagcgagc cagaggacct
     2881 tgagaatggg gaagagagct ctgctgagct ggattaccaa gacccagatg ctacttccct
     2941 caagtacgtt agtggtgatg tcacccaccc tcaggctggg gccgaggatg ctctcattgt
     3001 gcactgcgta gatgactgtg gccactgggg cagaggtggt ttatttacag ctctggaaaa
     3061 gcgatccgct gagccaagaa aaatatatga gctggctggg aaaatgaaag acctgagttt
     3121 gggaggtgtc cttttatttc ctgttgatga taaagaatca agaaacaaag ggcaagattt
     3181 gttggccttg attgtggctc agcatcgtga tcgttccaat gtcctgtctg gcattaagat
     3241 ggcagcccta gaagagggcc tgaagaagat atttttagca gcaaaaaaga agaaagcaag
     3301 tgttcatctt ccacgtattg gacatgccac gaaaggtttt aactggtatg gtactgagcg
     3361 acttattcgg aaacatctgg ctgcaagagg catcccaact tacatatatt attttcctag
     3421 aagcaagtct gctgtccttc attcacagtc ttcatcttcc tcctcaagac agctggtgcc
     3481 ttaaggatcc gtcgagcacc accaccacca ccaccaccac taattgatta atacctaggc
     3541 tgctaaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     3601 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     3661 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3721 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3781 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3841 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3901 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3961 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4021 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4081 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     4141 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4201 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     4261 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     4321 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     4381 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     4441 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     4501 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     4561 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     4621 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     4681 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     4741 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     4801 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     4861 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     4921 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     4981 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     5041 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     5101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5281 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5761 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5821 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5881 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5941 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     6001 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     6061 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     6121 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     6181 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     6241 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     6301 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     6361 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     6421 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     6481 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     6541 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     6601 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     6661 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     6721 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     6781 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgct agtcatgccc
     6841 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     6901 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     6961 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7021 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     7081 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     7141 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     7201 atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc
     7261 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     7321 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     7381 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     7441 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     7501 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     7561 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     7621 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     7681 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     7741 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     7801 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     7861 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     7921 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     7981 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     8041 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     8101 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     8161 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     8221 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     8281 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     8341 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatcgatc
     8401 tcgatcccgc gaaat
//
