LOCUS       sequence-334563-        8310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses codon optimised full length His tagged PARP1 in bacterial 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8310)
  AUTHORS   Ogden TEH, Yang JC, Schimpl M, Easton LE, Underwood E, Rawlins PB, 
            McCauley MM, Langelier MF, Pascal JM, Embrey KJ, Neuhaus D
  TITLE     Dynamics of the HD regulatory subdomain of PARP-1; substrate access 
            and allostery in PARP activation and inhibition.
  JOURNAL   Nucleic Acids Res. 2021 Feb 26;49(4):2266-2288. doi: 
            10.1093/nar/gkab020.
  PUBMED    33511412
REFERENCE   2  (bases 1 to 8310)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2021-02-26- 26"; volume: "49"; issue: 
            "4"; pages: "2266-2288"
COMMENT     Sequence Label: PARP1 FL
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     111..130
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     214..233
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /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             272..294
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             308..325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3388..3405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3472..3519
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3556..4011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3643..3662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3853..3874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4103..4918)
                     /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(4103..4918)
                     /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     4826..4845
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4910..4929
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5040..5628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5529..5548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5782..5799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5814..5953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5879..5898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6039..6061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6055..6246)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7055..8137)
                     /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     8099..8118
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8138..8215)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
       61 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
      121 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      181 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca
      301 tatgaaacac catcaccatc accatatgaa acaggcagaa agcagcgata aactgtatcg
      361 tgtggaatat gcaaaaagcg gtcgtgcaag ctgtaaaaaa tgtagcgaga gcattccgaa
      421 agatagcctg cgtatggcaa ttatggttca gagcccgatg tttgatggta aagtgccgca
      481 ttggtatcat ttcagctgct tttggaaagt tggtcatagc attcgtcatc cggatgttga
      541 agttgatggt tttagcgaac tgcgttggga tgatcagcag aaagtgaaaa aaaccgctga
      601 agccggtggt gttactggta aaggtcagga tggtattggt agcaaagcag aaaaaaccct
      661 gggtgatttt gcagcagaat atgccaaaag caatcgtagc acctgtaaag gctgcatgga
      721 aaaaattgaa aaaggtcagg tgcgtctgag caaaaaaatg gttgatccgg aaaaaccgca
      781 gctgggtatg attgatcgtt ggtatcatcc gggttgcttt gttaaaaatc gcgaagaact
      841 gggttttcgt ccggaatata gcgcaagcca gctgaaaggt tttagcctgc tggcaaccga
      901 agataaagaa gccctgaaaa aacagctgcc tggtgttaaa agcgaaggta aacgcaaagg
      961 tgatgaagtg gatggtgttg atgaggttgc caaaaaaaaa agcaaaaaag agaaagataa
     1021 agatagcaaa ctggaaaaag cactgaaagc ccagaatgat ctgatctgga acatcaaaga
     1081 cgaactgaaa aaagtgtgta gcaccaacga tctgaaagaa ctgctgatct ttaataaaca
     1141 gcaggttccg agcggtgaaa gcgcaattct ggatcgtgtt gccgatggta tggtttttgg
     1201 tgcactgctg ccgtgtgaag aatgtagcgg tcagctggtt tttaaaagcg acgcatatta
     1261 ttgtaccggt gatgttaccg catggaccaa atgtatggtt aaaacccaga ccccgaatcg
     1321 taaagaatgg gtgaccccga aagaattccg tgagattagc tatctgaaaa aactgaaagt
     1381 taaaaaacag gatcgcattt ttccgcctga aaccagcgca agcgttgcag caacaccgcc
     1441 tccgagcacc gcatctgcac cggcagcagt taatagcagc gcaagcgcag ataaaccgct
     1501 gagcaacatg aaaattctga ccctgggtaa actgagccgt aacaaagatg aagtgaaagc
     1561 catgattgaa aaactgggtg gtaaactgac cggcaccgca aataaagcaa gcctgtgcat
     1621 cagcaccaaa aaagaggtgg aaaaaatgaa taaaaaaatg gaagaagtga aagaagccaa
     1681 tattcgcgtt gttagcgaag attttctgca agatgttagc gcaagcacca aaagcctgca
     1741 agaactgttt ctggcacata ttctgagccc gtggggtgca gaagttaaag cagaaccggt
     1801 tgaagttgtt gcaccgcgtg gtaaaagcgg tgcagcactg tctaaaaaat ccaaaggcca
     1861 ggtgaaagaa gaaggcatca acaaaagcga aaaacgtatg aaactgaccc tgaaaggtgg
     1921 tgcagcagtt gatccggata gcggtctgga acatagcgca catgttctgg aaaaaggtgg
     1981 taaagttttt agcgcaaccc tgggtctggt tgatattgtt aaaggcacca atagctacta
     2041 taaactgcaa ctgctggaag atgataaaga aaaccgctat tggatttttc gtagctgggg
     2101 tcgtgttggc accgttattg gcagcaataa actggaacaa atgccgagca aagaagatgc
     2161 catcgaacac ttcatgaaac tgtatgaaga aaaaaccggt aatgcctggc atagcaaaaa
     2221 cttcaccaaa tatccgaaaa aattctatcc gctggaaatc gattatggtc aggatgaaga
     2281 agcagttaaa aaactgaccg tgaatccggg tacaaaaagc aaactgccga aaccggttca
     2341 ggatctgatc aaaatgatct ttgatgtgga aagcatgaaa aaagccatgg tggaatatga
     2401 aattgacctg caaaaaatgc cgctgggcaa actgagcaaa cgtcagattc aggcagcata
     2461 tagcattctg agcgaagttc agcaggcagt tagccagggt agcagcgata gccagattct
     2521 ggatctgagc aatcgctttt ataccctgat tccgcacgat tttggtatga aaaaaccgcc
     2581 tctgctgaat aatgcagata gcgttcaggc aaaagcagaa atgctggata acctgctgga
     2641 tattgaagtt gcatatagcc tgctgcgtgg tggttctgat gatagcagca aagatccgat
     2701 cgatgtgaac tatgagaaac tgaaaaccga tattaaagtg gttgatcgtg atagcgaaga
     2761 agccgaaatt atccgcaaat acgtgaaaaa tacccatgca accacccata atgcctatga
     2821 tctggaagtg attgatatct ttaaaatcga acgtgaaggt gaatgtcagc gctataaacc
     2881 gtttaaacag ctgcataatc gtcgtctgct gtggcatggt agccgtacca ccaattttgc
     2941 aggtattctg agccagggtc tgcgtattgc accgcctgaa gcaccggtta ccggttatat
     3001 gtttggcaaa ggcatctatt ttgccgatat ggttagcaaa agcgccaatt attgtcatac
     3061 cagccagggt gatccgattg gtctgattct gctgggtgaa gttgcactgg gtaacatgta
     3121 tgaactgaaa catgccagcc atattagcaa actgcctaaa ggcaaacata gcgttaaagg
     3181 tctgggcaaa accacaccgg atccgagcgc aaatattagc ctggatggtg tggatgttcc
     3241 tctgggcacc ggtattagca gcggtgttaa tgataccagc ctgctgtata acgaatacat
     3301 cgtgtatgat attgcccagg tgaatctgaa atacctgctg aaactgaaat tcaacttcaa
     3361 aaccagcctg tggtagtaag actcgagcac caccaccacc accactgaga tccggctgct
     3421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3481 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3541 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3601 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3661 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3721 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3781 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3841 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3901 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3961 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     4021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4081 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     4141 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     4201 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     4261 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     4321 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     4381 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     4441 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     4501 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4561 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4621 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     4681 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     4741 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4801 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     4861 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     4921 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     4981 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5041 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5101 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5161 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5221 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5281 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5341 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5401 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5461 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5521 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5581 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5641 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5701 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5761 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     5821 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     5881 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     5941 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6061 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     6121 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     6181 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     6241 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     6301 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     6361 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     6421 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     6481 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     6541 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     6601 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     6661 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     6721 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     6781 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     6841 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     6901 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     6961 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     7021 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7141 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     7201 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     7261 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     7321 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     7381 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     7441 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     7501 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     7561 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     7621 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     7681 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     7741 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     7801 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     7861 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     7921 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     7981 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     8041 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     8101 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     8161 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     8221 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     8281 ccgttgagca ccgccgccgc aaggaatggt
//
