LOCUS       sequence-434366-        6780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for E. coli producing PARP2 catalytic 
            protein (without Regulatory domain); with N-terminal CFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6780)
  AUTHORS   Dhakar SS, Galera-Prat A, Lehtio L
  TITLE     High-throughput screening assay for PARP-HPF1 interaction inhibitors
            to affect DNA damage repair.
  JOURNAL   Sci Rep. 2024 Feb 16;14(1):3875. doi: 10.1038/s41598-024-54123-8.
  PUBMED    38365924
REFERENCE   2  (bases 1 to 6780)
  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: "Sci Rep."; 
            date: "2024-02-16"; volume: "14(1):3875. doi"; pages: " 
            10.1038/s41598-024-54123-8"
COMMENT     Sequence Label: PARP2-c098
FEATURES             Location/Qualifiers
     source          1..6780
                     /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             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(110..112,113..823)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(152..173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(413..432)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     760..781
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             848..868
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1645..1662
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1715..1733)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1729..1776
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1813..2268
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1900..1919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2110..2131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2360..3175)
                     /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(2360..3175)
                     /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     3083..3102
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3167..3186
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3297..3885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3786..3805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4039..4056
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4071..4210
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4136..4155)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4296..4318
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4312..4503)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5312..6394)
                     /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     6356..6375
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6395..6472)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6678..6697
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatg tgagcaaggg
      121 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      181 ccacaggttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      241 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      301 gacctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
      361 caagtccgcc atgcccgaag gctacgtcca ggagcgtacc atcttcttca aggacgacgg
      421 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      481 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      541 cgccatcagc gacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccca
      601 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      661 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
      721 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      781 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtcttctg gtgtagatct
      841 gggtaccgag aacttgtact tccaatccat gctgcagtct ccggaacatc cgctggacca
      901 acactaccgt aacctgcatt gtgcgctgcg cccgctggat cacgaatcat acgaattcaa
      961 ggtgatctca cagtacctgc aatcgaccca tgccccgacg cactcggatt ataccatgac
     1021 gctgctggac ctgtttgaag ttgaaaaaga tggtgaaaag gaagccttcc gtgaagatct
     1081 gcacaatcgc atgctgctgt ggcatggcag ccgtatgtct aactgggtgg gtatcctgag
     1141 ccatggtctg cgtattgcac cgccggaagc tccgatcacc ggctacatgt ttggcaaagg
     1201 tatttatttc gcagatatga gttccaaaag cgcaaattac tgctttgctt ctcgtctgaa
     1261 aaacacgggt ctgctgctgc tgagtgaagt tgctctgggc cagtgtaatg aactgctgga
     1321 agcgaacccg aaagccgaag gtctgctgca gggcaaacac tccaccaagg gcctgggtaa
     1381 aatggcaccg tcatcggctc atttcgtcac cctgaatggt agtacggtgc cgctgggtcc
     1441 ggcatccgac accggtattc tgaatccgga tggctatacg ctgaattata acgaatacat
     1501 cgtgtataac ccgaaccagg tccgtatgcg ttatctgctg aaggtccaat tcaactttct
     1561 gcaactgtgg taagacggtc tccagtaaag gtggatacgg atccgaattc gagctccgtc
     1621 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
     1681 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1741 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1801 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1861 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1921 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1981 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2041 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2101 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2161 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2221 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2281 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2341 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     2401 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     2461 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     2521 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     2581 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     2641 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     2701 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     2761 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     2821 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     2881 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     2941 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     3001 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     3061 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     3121 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
     3181 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
     3241 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3301 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3361 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3421 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3481 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3541 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3601 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3661 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3721 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3781 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3841 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3901 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3961 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4021 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4081 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
     4141 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4201 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4261 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4321 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4381 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4441 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4501 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     4561 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4621 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4681 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     4741 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     4801 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     4861 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     4921 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     4981 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     5041 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     5101 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     5161 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     5221 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     5281 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5341 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5401 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     5461 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     5521 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     5581 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     5641 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     5701 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     5761 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     5821 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     5881 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     5941 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     6001 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     6061 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     6121 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     6181 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     6241 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     6301 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     6361 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     6421 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     6481 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     6541 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     6601 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     6661 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     6721 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
