LOCUS       sequence-204960-        6072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression in E.coli and affinity purification of N-terminally 
            GST-tagged human cGAS amino acid 157-522.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6072)
  AUTHORS   Mackenzie KJ, Carroll P, Martin CA, Murina O, Fluteau A, Simpson DJ,
            Olova N, Sutcliffe H, Rainger JK, Leitch A, Osborn RT, Wheeler AP, 
            Nowotny M, Gilbert N, Chandra T, Reijns MAM, Jackson AP
  TITLE     cGAS surveillance of micronuclei links genome instability to innate 
            immunity.
  JOURNAL   Nature. 2017 Aug 24;548(7668):461-465. doi: 10.1038/nature23449. 
            Epub 2017 Jul 24.
  PUBMED    28738408
REFERENCE   2  (bases 1 to 6072)
  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"; doi: 
            "10.1038/nature23449"; journalName: "Nature"; date: "2017-08-24- 
            24"; volume: "548"; issue: "7668"; pages: "461-465"
COMMENT     Sequence Label: pGEX6P1-hcGAS(157-522)
FEATURES             Location/Qualifiers
     source          1..6072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2122..2144)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2289..2307)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2375..2479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2480..2548,2549..3340)
                     /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(2698..2717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3511..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4000..4019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4253..4270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4343..4420
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4421..5503
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4440..4459)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5516..5537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5552..5557,5558..5575,5576..5582)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5590..5606
                     /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     5614..5630
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5614..5630
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5626..6072,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5646..5663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5646..5662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5655..5677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc gatgcggcgc
      961 ctggggcctc gaagctccgg gcggttttgg agaagttgaa gctcagccgc gatgatatct
     1021 ccacggcggc ggggatggtg aaaggggttg tggaccacct gctgctcaga ctgaagtgcg
     1081 actccgcgtt cagaggcgtc gggctgctga acaccgggag ctactatgag cacgtgaaga
     1141 tttctgcacc taatgaattt gatgtcatgt ttaaactgga agtccccaga attcaactag
     1201 aagaatattc caacactcgt gcatattact ttgtgaaatt taaaagaaat ccgaaagaaa
     1261 atcctctgag tcagttttta gaaggtgaaa tattatcagc ttctaagatg ctgtcaaagt
     1321 ttaggaaaat cattaaggaa gaaattaacg acattaaaga tacagatgtc atcatgaaga
     1381 ggaaaagagg agggagccct gctgtaacac ttcttattag tgaaaaaata tctgtggata
     1441 taaccctggc tttggaatca aaaagtagct ggcctgctag cacccaagaa ggcctgcgca
     1501 ttcaaaactg gctttcagca aaagttagga agcaactacg actaaagcca ttttaccttg
     1561 tacccaagca tgcaaaggaa ggaaatggtt tccaagaaga aacatggcgg ctatccttct
     1621 ctcacatcga aaaggaaatt ttgaacaatc atggaaaatc taaaacgtgc tgtgaaaaca
     1681 aagaagagaa atgttgcagg aaagattgtt taaaactaat gaaatacctt ttagaacagc
     1741 tgaaagaaag gtttaaagac aaaaaacatc tggataaatt ctcttcttat catgtgaaaa
     1801 ctgccttctt tcacgtatgt acccagaacc ctcaagacag tcagtgggac cgcaaagacc
     1861 tgggcctctg ctttgataac tgcgtgacat actttcttca gtgcctcagg acagaaaaac
     1921 ttgagaatta ttttattcct gaattcaatc tattctctag caacttaatt gacaaaagaa
     1981 gtaaggaatt tctgacaaag caaattgaat atgaaagaaa caatgagttt ccagtttttg
     2041 atgaattttg agtcgactcg agcggccgca tcgtgactga ctgacgatct gcctcgcgcg
     2101 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2161 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2221 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata attcttgaag
     2281 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     2341 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2401 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2461 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2581 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2641 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2701 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     2761 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2881 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3001 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3061 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4201 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4261 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4321 tatttcacac cgcataaatt ccgacaccat cgaatggtgc aaaacctttc gcggtatggc
     4381 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     4441 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     4501 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     4561 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     4621 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     4681 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     4741 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     4801 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     4861 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     4921 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     4981 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     5041 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     5101 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     5161 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     5221 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgtcaac
     5281 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     5341 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     5401 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5461 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5521 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     5581 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     5641 gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5701 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5761 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg
     5821 gcaccagaag cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc
     5881 gtcgtcccct caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtaacc
     5941 tatcccatta cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg
     6001 ctcacattta atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat
     6061 ggcgttggaa tt
//
