LOCUS       sequence-379671-        8334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PARP1 with an N-terminal His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8334)
  AUTHORS   Langelier MF, Servent KM, Rogers EE, Pascal JM
  TITLE     A third zinc-binding domain of human poly(ADP-ribose) polymerase-1 
            coordinates DNA-dependent enzyme activation.
  JOURNAL   J Biol Chem. 2008 Feb 15;283(7):4105-14. doi: 
            10.1074/jbc.M708558200. Epub 2007 Nov 30.
  PUBMED    18055453
REFERENCE   2  (bases 1 to 8334)
  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: 
            "10.1074/jbc.M708558200"; journalName: "J Biol Chem"; date: 
            "2008-02-15- 15"; volume: "283"; issue: "7"; pages: "4105-14"
COMMENT     Sequence Label: pET 28 full-length human PARP1
FEATURES             Location/Qualifiers
     source          1..8334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3212..3229)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(3239..3256)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3275..3297
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3312..3336
                     /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)."
     primer_bind     complement(3336..3355)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3337..3355)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3439..3458)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3664..3741
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3742..4824
                     /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     complement(3761..3780)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5633..5824
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5818..5840)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5926..6065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5981..6000
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6080..6097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6251..6839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6331..6350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6950..6969)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6961..7776
                     /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             6961..7776
                     /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     complement(7034..7053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(7868..8323)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8005..8026)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8217..8236
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttaccac agggaggtct
      181 taaaattgaa tttcagtttc agcagatact tcagatttac ctgagcaata tcatagacaa
      241 tgtactcgtt atatagtaga gaggtgtcat tcacaccaga tgaaatcccg gtcccaagag
      301 gaacgtctac accatccaga ctaatgttag ctgaaggatc aggggtagtt ttgcccaaac
      361 ctttgacact gtgcttgccc ttgggtaact tgctgatatg tgaagcgtgc ttcagttcat
      421 acatgtttcc aagggcaact tctcccaaca ggattaagcc tattgggtct ccctgagacg
      481 tatggcagta gttggcactc ttggagacca tgtcagcgaa atagatccct ttaccaaaca
      541 tgtagcctgt cacgggcgct tcaggcgggg ctatccgaag accctgggac aggatcccag
      601 caaagttggt ggtcctggac ccgtgccaca gcaatcttcg gttatgaagc tgcttaaagg
      661 gcttgtaacg ctggcattcg ccttcacgct ctatcttaaa gatatcgatg acttccaagt
      721 catacgcatt gtgtgtggtt gcatgagtgt tcttaacata cttcctgatg atctcggctt
      781 cttcagaatc tctgtcaacc accttaatgt cagttttgag cttctcatag ttgacatcga
      841 tgggatcctt gctgctatca tcagaccctc ccctgagcag actgtaggcc acctcgatgt
      901 ccagcaggtt gtcaagcatt tccgccttgg cctgcacact gtctgcattg ttcaggagcg
      961 gaggcttctt catcccaaag tcgtggggga tcagggtgta aaagcgattt gagagatcca
     1021 ggatctgaga gtcgctgctg ccctgagaca ccgcctgctg gacctcactg aggatggagt
     1081 atgcggcctg gatctgcctt ttgctcagct tccccaaggg catcttctga aggtcgatct
     1141 catactccac catggctttc ttcatacttt ccacatcaaa gatcatcttg atgaggtcct
     1201 gaactggctt ggggagcttg gacttggtgc caggatttac tgtcagcttc ttcactgcct
     1261 cttcatcctg gccatagtca atctccaggg ggtagaactt tttgggatac ttcgtgaaat
     1321 ttttggagtg ccaagcgttc ccggtttttt cttcatataa tttcatgaag tgctcaatgg
     1381 catcctcctt ggacggcatc tgttccagtt tgttgctacc gatcaccgta cccacacggc
     1441 cccaggacct gaatatccaa tacctgtttt ccttgtcgtc ctccagaagc tggagcttgt
     1501 agtaggagtt ggttccttta acgatgtcca ccaggccaag ggtggcactg aagaccttcc
     1561 cacctttctc caggacatgc gcagagtgtt ccagtccaga atcaggatcc acagctgctc
     1621 ctcctttaag agttaatttc attctctttt cagatttgtt gataccttcc tccttgacct
     1681 ggcccttgct ttttttggag agcgcagccc ctgacttccc tcttggggcc acaacttcaa
     1741 caggctctgc cttcacctct gccccccaag gggacaagat gtgcgctaag aacaactcct
     1801 gaaggctctt ggtggaggcg gagacgtcct ggaggaagtc ctcagacaca actcggatgt
     1861 tggcttcctt tacttcctcc atcttcttat tcatcttttc cacctccttt ttggtgctga
     1921 tgcacaggga agccttgttg gccgtccccg tcaacttccc cccgagtttc tcaatcatgg
     1981 ccttcacttc atccttgttc cgggacagct tcccgagagt caggatcttc atgttggata
     2041 atggcttatc tgctgaagca gaggagttca cagcagcagg agccgaggct gtggagggcg
     2101 gaggcgtggc cgccacggag gcgctggttt ctggggggaa tatacggtcc tgctttttaa
     2161 ccttcaattt cttgaggtaa gagatttctc ggaattcctt tggggttacc cactccttcc
     2221 ggttgggtgt ctgtgtcttg accatacact tggtccaggc agtgacgtcc ccagtgcagt
     2281 aataggcatc gctcttgaag accagctgac ccgagcattc ctcgcaggga aggagggcac
     2341 cgaacaccat gccatcagct actcggtcca agatcgccga ctccccagaa ggcacttgct
     2401 gcttgttgaa gatgagtagc tccttcaggt cattagttga acacactttc tttagctcgt
     2461 ccttgatgtt ccagatcagg tcgttctgag cctttagggc tttttcaagc ttactatcct
     2521 tgtctttttc ttttttagat ttcttcttcg ccacttcatc cactccatcc acctcatcgc
     2581 cttttctctt tccttcactc ttgactcctg ggagctgctt cttcagggct tccttatcct
     2641 ctgtagcaag gaggctgaag cccttgagct gactcgcact gtactcgggc cggaaaccca
     2701 gctcctccct gttcttgaca aagcagcctg gatggtacca gcggtcaatc atgcctagct
     2761 gtggcttctc cgggtccacc atcttcttgg acaggcgcac ctggcccttt tctatcttct
     2821 ccatacaccc cttgcacgta cttctgttgg acttggcata ctctgctgca aagtcaccca
     2881 gagtcttctc tgccttgcta ccaattccat cctggccttt gcctgtcact cctccagctt
     2941 ccgctgtctt cttgactttc tgctggtcat cccaccgaag ctcagagaac ccatccacct
     3001 caacgtcagg gtgccggatg gagtggccca ccttccagaa gcaggagaag tggtaccagt
     3061 gtgggacttt tccatcaaac atgggcgact gcaccatgat ggccatccgg agcgagtcct
     3121 tggggatgct ctcgctgcat ttcttgcaag aggcgcgccc gctcttggcg tactcgactc
     3181 gatacagctt atcagaagac tccgccatat ggctgccgcg cggcaccagg ccgctgctgt
     3241 gatgatgatg atgatggctg ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt
     3301 atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc
     3361 gcgggatcga gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     3421 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     3481 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     3541 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     3601 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
     3661 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     3721 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     3781 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     3841 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     3901 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     3961 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     4021 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     4081 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     4141 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     4201 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     4261 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     4321 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     4381 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     4441 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     4501 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     4561 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     4621 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     4681 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     4741 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4801 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     4861 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     4921 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     4981 aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc gctggagcgc
     5041 gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt
     5101 cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg ccggcatggc
     5161 ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     5221 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     5281 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     5341 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     5401 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     5461 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     5521 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     5581 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     5641 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     5701 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     5761 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     5821 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5881 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     5941 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     6001 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6061 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6661 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6901 ggattttggt catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     6961 atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat
     7021 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     7081 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     7141 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     7201 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     7261 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     7321 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     7381 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     7441 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     7501 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     7561 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     7621 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     7681 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     7741 ttgcagtttc atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat
     7801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7861 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     7921 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     7981 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     8041 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     8101 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     8161 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     8221 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     8281 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gcca
//
