LOCUS       sequence-434367-        7116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for E. coli producing full length HPF1 
            protein with N-terminal YFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7116)
  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 7116)
  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: HPF1-c004
FEATURES             Location/Qualifiers
     source          1..7116
                     /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 YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     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             1981..1998
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2051..2069)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2065..2112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2149..2604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2236..2255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2446..2467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2696..3511)
                     /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(2696..3511)
                     /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     3419..3438
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3503..3522
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3633..4221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4122..4141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4375..4392
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4407..4546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4472..4491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4632..4654
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4648..4839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5648..6730)
                     /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     6692..6711
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6731..6808)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7014..7033
                     /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 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      241 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
      301 cggctacggc ctgatgtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
      361 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      421 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      481 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      541 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
      601 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      661 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
      721 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
      781 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtcttctg gtgtagatct
      841 gggtaccgag aacttgtact tccaatccat ggtgggtggc ggtggcaaac gtcgtccggg
      901 tggcgaaggt ccgcagtgcg agaagaccac cgacgttaag aaaagcaaat tttgcgaagc
      961 ggacgtgagc agcgatctgc gtaaagaggt ggaaaaccac tacaaactga gcctgccgga
     1021 agatttctat cacttttgga agttctgcga ggaactggac ccggagaaac cgagcgatag
     1081 cctgagcgcg agcctgggtc tgcaactggt gggcccgtac gacatcctgg cgggcaagca
     1141 caaaaccaag aaaaagagca ccggcctgaa ctttaacctg cactggcgtt tctactatga
     1201 cccgccggag ttccagacca tcattatcgg tgataacaag acccaatacc acatgggcta
     1261 ttttcgtgac agcccggatg agttcccggt gtatgttggt attaacgagg cgaaaaagaa
     1321 ctgcattatc gtgccgaacg gcgacaacgt gtttgcggcg gttaaactgt tcctgaccaa
     1381 aaagctgcgt gaaattaccg acaaaaagaa aatcaacctg ctgaagaaca ttgatgagaa
     1441 actgaccgaa gcggcgcgtg agctgggtta cagcctggag cagcgtaccg ttaagatgaa
     1501 acaacgtgat aagaaagtgg ttaccaagac ctttcacggt gcgggcctgg tggttccggt
     1561 ggacaaaaac gatgttggct atcgtgagct gccggaaacc gacgcggatc tgaagcgtat
     1621 ttgcaaaacc atcgtggaag cggcgagcga cgaggaacgt ctgaaagcgt tcgcgccgat
     1681 ccaggagatg atgacctttg ttcaattcgc gaacgacgaa tgcgattacg gtatgggcct
     1741 ggagctgggt atggacctgt tctgctacgg cagccactat ttccacaagg ttgcgggtca
     1801 gctgctgccg ctggcgtata acctgctgaa acgtaacctg tttgcggaaa ttatcgagga
     1861 acacctggcg aaccgtagcc aggagaacat tgatcaactg gcggcgtaag acggtctcca
     1921 gtaaaggtgg atacggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag
     1981 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     2041 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     2101 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     2161 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2221 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2281 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2341 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2401 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2461 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2521 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2581 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2641 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     2701 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     2761 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     2821 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     2881 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     2941 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3001 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     3061 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     3121 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     3181 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3241 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3301 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3361 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     3421 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     3481 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     3541 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3601 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3661 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3721 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3781 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3841 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3901 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3961 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4021 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4081 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4141 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4201 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4261 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4321 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4381 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     4441 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4501 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4561 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4621 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4681 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4741 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4801 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     4861 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4921 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4981 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     5041 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     5101 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5161 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5221 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     5281 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     5341 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     5401 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     5461 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     5521 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     5581 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     5641 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5701 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     5761 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     5821 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     5881 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     5941 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     6001 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     6061 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     6121 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     6181 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     6241 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     6301 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     6361 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     6421 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     6481 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     6541 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     6601 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     6661 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     6721 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     6781 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     6841 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     6901 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     6961 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     7021 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     7081 ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
