LOCUS       sequence-259515-        6547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mouse PARG(439-959) in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6547)
  AUTHORS   Wang Z, Gagne JP, Poirier GG, Xu W
  TITLE     Crystallographic and biochemical analysis of the mouse 
            poly(ADP-ribose) glycohydrolase.
  JOURNAL   PLoS One. 2014 Jan 21;9(1):e86010. doi: 
            10.1371/journal.pone.0086010. eCollection 2014.
  PUBMED    24465839
REFERENCE   2  (bases 1 to 6547)
  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: "PLoS One.";
            date: "2014-01-21"; volume: "9(1):e86010. doi"; pages: " 
            10.1371/journal.pone.0086010. eCollection 2014"
COMMENT     Sequence Label: pGEX-4T1 mouse PARG (439-959)
FEATURES             Location/Qualifiers
     source          1..6547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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)."
     CDS             40..693
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     651..673
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             700..717
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             727..747
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2379..2401)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2546..2564)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2632..2736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2737..2805,2806..3597)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2955..2974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3768..4356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4257..4276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4510..4527
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4600..4677
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4678..5760
                     /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(4697..4716)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5809..5814,5815..5832,5833..5839)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5847..5863
                     /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     5871..5887
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5871..5887
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5883..6410
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5903..5920)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5903..5919)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5912..5934)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(6512..6517,6518..6533,6534..6540)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat actaggttat
       61 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
      121 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
      181 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
      241 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
      301 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
      361 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
      421 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
      481 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
      541 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
      601 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
      661 gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggttccgcg tggatccccg
      721 gaattcgaaa acctgtattt tcagggcaac ctccctccag agaagaagtg gctgggaact
      781 cctattgagg aaatgagaaa aatgcctcgg tgtgggatcc atttgccttc cttaagacca
      841 tctgcaagtc acacagtgac tgttcgggta gaccttctga gagcaggaga ggttccgaaa
      901 ccttttccaa cacattacaa agatttgtgg gataacaaac atgtgaaaat gccttgttcg
      961 gaacaaaact tgtaccctgt ggaagatgag aatggtgagc gaactgcagg gagtaggtgg
     1021 gagctcattc agactgcact tctcaacaaa ttcacacgac cccagaactt gaaggatgcg
     1081 attctgaaat acaatgtggc atattctaag aaatgggact ttacagcttt ggttgatttc
     1141 tgggataagg tacttgaaga agcagaggcc caacatttat atcagtccat tttacctgac
     1201 atggtgaaaa ttgcactctg tctgccaaat atttgcaccc agccaatacc actcctgaaa
     1261 cagaagatga atcattctgt cacgatgtca caggaacaga tcgccagtct tttagctaat
     1321 gctttcttct gcacatttcc ccgacggaat gccaagatga aatcggagta ttctagttac
     1381 ccagacatta acttcaatcg gttgtttgaa ggacgttcat caaggaaacc agaaaaactg
     1441 aaaacactct tctgctactt tcgaagagtc acagagaaaa aacctacagg attggtgaca
     1501 tttacaagac agagtcttga agattttcca gaatgggaaa ggtgtgaaaa gcctctgaca
     1561 cgcttacacg tcacttacga gggtaccata gaaggcaacg gccgaggcat gctacaggtg
     1621 gattttgcaa atcgttttgt tggaggtggt gtgactggtg cgggacttgt acaagaagaa
     1681 atcagatttt taatcaatcc tgaattgatt gtttcacggc tgttcactga ggtgctggat
     1741 cacaatgagt gtcttattat cacaggtact gaacagtaca gtgaatacac aggctatgct
     1801 gaaacttatc gttgggcccg aagccatgaa gatgggagtg aaaaggacga ttggcagcgg
     1861 cgctgcacgg agatcgttgc cattgacgca cttcacttca gacgctacct cgatcagttt
     1921 gtgcctgaga aagtgagacg tgagcttaac aaggcttact gcggattcct ccgtcctgga
     1981 gttccttctg aaaatctttc tgcagtggcc acgggaaact ggggctgtgg tgcctttggg
     2041 ggtgacgcta gattaaaagc cttaatacag atcctggcag ctgctgcggc tgaacgtgac
     2101 gtggtttatt tcacctttgg ggactcagag ttgatgagag acatttacag catgcacact
     2161 ttccttaccg agaggaagct ggatgttgga aaagtgtaca agttattgct tagatactac
     2221 aatgaagaat gcagaaactg ttccacccct ggaccagaca tcaagcttta tccattcata
     2281 taccatgctg ttgagtcaag tgcagagacc tgactcgagc ggccgcatcg tgactgactg
     2341 acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     2401 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     2461 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     2521 gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2581 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2641 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2701 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2761 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2821 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2881 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2941 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     3001 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3061 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3121 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3181 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3241 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     3301 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3361 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3421 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3481 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3541 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3601 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3661 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3721 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3781 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3841 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3901 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3961 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4021 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4081 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4141 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4201 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4261 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4321 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4381 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4441 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4501 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4561 cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggcgcaaa
     4621 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     4681 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     4741 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     4801 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     4861 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     4921 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     4981 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     5041 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     5101 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     5161 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     5221 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     5281 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     5341 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     5401 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     5461 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     5521 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     5581 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     5641 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     5701 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5761 gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt tacactttat
     5821 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5881 ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     5941 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     6001 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     6061 gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc
     6121 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca
     6181 tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc
     6241 cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga
     6301 cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc accaatgctt
     6361 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     6421 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     6481 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
     6541 tgtggaa
//
