LOCUS       sequence-216916-        6744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ARH3 (amino acids 14-363) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  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 6744)
  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-ARH3-His-Sumo-HA
FEATURES             Location/Qualifiers
     source          1..6744
                     /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             4428..4454
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4428..4454
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4504..4521
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4574..4592)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4588..4635
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4672..5127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4759..4778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4969..4990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5219..6034)
                     /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(5219..6034)
                     /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     5942..5961
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6026..6045
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6156..6744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6645..6664
                     /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 tggtgctgga gcggcacgta
     3361 gcctgagccg cttccgtggc tgcttggctg gtgctctgtt aggagattgc gtagggtcct
     3421 tttacgaagc tcatgacacc gtcgatttaa cgagcgtact tcgtcacgtt caaagcctgg
     3481 aacctgatcc cgggacgcct ggttctgagc gtactgaagc attatattac accgacgata
     3541 ctgccatggc ccgcgccctt gttcagtctc ttttagctaa ggaggcgttc gacgaggtgg
     3601 acatggctca ccgtttcgcg caggaataca agaaagaccc cgaccgtgga tacggtgcag
     3661 gtgtagtgac cgtatttaag aaacttttaa acccgaaatg tcgtgacgtt tttgaaccag
     3721 cccgtgctca attcaatggt aaaggttcgt atggcaacgg tggggctatg cgtgtcgcag
     3781 gcatctcact ggcgtactct tcggttcaag atgtccagaa gtttgcccgt ttatcagcgc
     3841 aattgacaca cgcttcaagc cttgggtaca acggagccat cttacaggca cttgccgtac
     3901 atttagcatt gcagggggag tcaagctcag agcatttctt gaaacaactg cttggccaca
     3961 tggaggattt ggaaggagat gcacaatccg tacttgacgc ccgtgaactg ggcatggaag
     4021 agcgtcccta ctcttcccgc cttaagaaga ttggagagtt gctggatcag gcttcggtca
     4081 ctcgcgagga agttgtatca gaattgggaa acgggatcgc tgcgtttgag tctgtaccaa
     4141 ctgccatcta ctgctttctg cgctgcatgg aaccggaccc agagatcccc tctgcgttca
     4201 actccctgca acgcacgctg atctatagca tttcactggg aggcgatacc gatactattg
     4261 ctaccatggc tggtgccatt gctggcgcct attatggaat ggatcaagtg ccggaaagct
     4321 ggcagcaaag ctgcgagggg tatgaggaga ctgacattct tgcgcagtcc ttacaccgtg
     4381 tttttcaaaa gtccgcgcag tccttacacc gtgtttttca aaagtcctac ccctacgacg
     4441 tgcccgacta cgccagatcc taagaattcg agctccgtcg acaagcttgc ggccgcactc
     4501 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag
     4561 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     4621 ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct
     4681 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4741 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4801 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4861 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4921 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4981 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5041 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5101 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     5161 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
     5221 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     5281 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     5341 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     5401 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     5461 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     5521 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     5581 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     5641 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     5701 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     5761 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     5821 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     5881 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     5941 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     6001 tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     6061 aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga
     6121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6301 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6721 tcaggggggc ggagcctatg gaaa
//
