LOCUS       sequence-270825-        6714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full length HPF1 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6714)
  AUTHORS   Liszczak G, Diehl KL, Dann GP, Muir TW
  TITLE     Acetylation blocks DNA damage-induced chromatin ADP-ribosylation.
  JOURNAL   Nat Chem Biol. 2018 Jul 16. pii: 10.1038/s41589-018-0097-1. doi: 
            10.1038/s41589-018-0097-1.
  PUBMED    30013063
REFERENCE   2  (bases 1 to 6714)
  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: "Nat Chem 
            Biol. 2018 Jul 16. pii: 10.1038/s41589-018-0097-1. doi: 
            10.1038/s41589-018-0097-1."
COMMENT     Sequence Label: pET30-HPF1-His-Sumo-Flag
FEATURES             Location/Qualifiers
     source          1..6714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    186..325
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(251..270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     411..433
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(427..618)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1427..2509)
                     /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     2471..2490
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2510..2587)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2793..2812
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2900..2919
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2900..2918
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2919..2943
                     /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             2958..2980
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3000..3017
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3027..3044
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3051..3344
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             3354..3377
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4474..4491
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4544..4562)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4558..4605
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4642..5097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4729..4748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4939..4960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5189..6004)
                     /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(5189..6004)
                     /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     5912..5931
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5996..6015
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6126..6714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6615..6634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
      241 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
      301 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
      361 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
      421 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
      481 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
      541 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
      601 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
      661 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
      721 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
      781 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
      841 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
      901 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
      961 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     1021 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     1081 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     1141 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     1201 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     1261 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     1321 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     1381 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     1441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1501 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     1561 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     1621 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     1681 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
     1741 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     1801 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     1861 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     1921 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     1981 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     2041 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     2101 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     2161 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     2221 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     2281 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     2341 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     2401 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     2461 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     2521 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     2581 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     2641 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     2701 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     2761 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     2821 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     2881 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
     2941 tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg ggcagcagcc
     3001 atcatcatca tcatcacggc agcggcctgg tgccgcgcgg cagcgctagc atgtcggact
     3061 cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct gagactcaca
     3121 tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa aagaccactc
     3181 ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg gactccttaa
     3241 gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat ttggacatgg
     3301 aggataacga tattattgag gctcacagag aacagattgg tggttcaggt agtgactaca
     3361 aagacgatga cgacaagggc ggatctggca tggtcggcgg tggcgggaag cgcaggcccg
     3421 gcggggaggg gccgcagtgt gaaaaaacaa ctgatgtgaa gaaaagtaaa ttctgtgaag
     3481 ctgatgtctc cagtgacctt cgaaaagaag tagaaaatca ttataagctt tctttacctg
     3541 aagatttcta tcacttctgg aagttctgtg aagaacttga tcctgaaaag ccatctgatt
     3601 cactttctgc aagccttgga cttcaattag ttggtcctta tgatatcctt gctggaaaac
     3661 ataaaacgaa gaaaaaatca acaggcctga attttaacct tcactggagg ttttactatg
     3721 atcctcctga gttccagacc attattattg gagataataa aactcagtac cacatggggt
     3781 atttcaggga ttctcctgat gaatttcctg tatatgttgg tataaatgaa gcaaagaaaa
     3841 attgtataat tgttccaaat ggagataatg tatttgctgc agtcaaatta tttttgacga
     3901 aaaaacttag agaaataacg gataaaaaga aaatcaatct cttgaaaaac atagatgaaa
     3961 aactcacaga agcagccaga gaattggggt actcgcttga acagagaacc gtgaagatga
     4021 aacagagaga taagaaagtt gtgacaaaga cctttcatgg tgcaggcttg gttgttccag
     4081 tagataaaaa tgatgttggg taccgagagc tccctgaaac agatgctgac ctcaagagaa
     4141 tttgcaagac aatagttgag gctgcaagtg atgaggagag actaaaagct tttgctccca
     4201 ttcaggaaat gatgactttt gtgcagtttg ctaatgatga atgtgattat ggcatggggc
     4261 ttgaattggg aatggatctc ttttgctatg gctcacatta ttttcataaa gttgctggcc
     4321 agcttttacc tcttgcatat aatctgttga agaggaatct gtttgcagaa attattgagg
     4381 agcatctggc aaacagaagt caagagaaca tagaccaact tgctgcatga taagaattcg
     4441 agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg
     4501 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     4561 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     4621 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4681 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4981 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5041 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5101 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5161 aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact
     5221 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     5281 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     5341 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     5401 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     5461 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     5521 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     5581 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     5641 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     5701 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     5761 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     5821 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     5881 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     5941 cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc
     6001 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa
     6061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6121 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6181 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6241 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6301 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6361 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6421 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6481 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6541 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6601 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6661 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaa
//
