LOCUS       sequence-245665-        5028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of truncated human cGAS protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5028)
  AUTHORS   Lama L, Adura C, Xie W, Tomita D, Kamei T, Kuryavyi V, Gogakos T, 
            Steinberg JI, Miller M, Ramos-Espiritu L, Asano Y, Hashizume S, Aida
            J, Imaeda T, Okamoto R, Jennings AJ, Michino M, Kuroita T, Stamford 
            A, Gao P, Meinke P, Glickman JF, Patel DJ, Tuschl T
  TITLE     Development of human cGAS-specific small-molecule inhibitors for 
            repression of dsDNA-triggered interferon expression.
  JOURNAL   Nat Commun. 2019 May 21;10(1):2261. doi: 10.1038/s41467-019-08620-4.
  PUBMED    31113940
REFERENCE   2  (bases 1 to 5028)
  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 
            Commun."; date: "2019-05-21"; volume: "10(1):2261. doi"; pages: " 
            10.1038/s41467-019-08620-4"
COMMENT     Sequence Label: pRSFDuet-sumo-h-cGASCD(K427E/K428E)
FEATURES             Location/Qualifiers
     source          1..5028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             110..127
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             140..433
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             1565..1609
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1647..1665)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1661..1708
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1938..2753)
                     /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(1938..2753)
                     /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     2661..2680
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2754..2845)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2861..3610)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2871..2888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3111..3130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3806..4888)
                     /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     4850..4869
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4889..4966)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5012..5028,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5012..5028,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg
      121 cggcagccac atggcaagca tgtcggactc agaagtcaat caagaagcta agccagaggt
      181 caagccagaa gtcaagcctg agactcacat caatttaaag gtgtccgatg gatcttcaga
      241 gatcttcttc aagatcaaaa agaccactcc tttaagaagg ctgatggaag cgttcgctaa
      301 aagacagggt aaggaaatgg actccttaag attcttgtac gacggtatta gaatccaagc
      361 tgatcagacc cctgaagatt tggacatgga ggataacgat attattgagg ctcacagaga
      421 acagattggt ggatccattc tcgtacggag ggatgcggcg cctggggcct cgaagctccg
      481 ggcggttttg gagaagttga agctcagccg cgatgatatc tccacggcgg cggggatggt
      541 gaaaggggtt gtggaccacc tgctgctcag actgaagtgc gactccgcgt tcagaggcgt
      601 cgggctgctg aacaccggga gctactatga gcacgtgaag atttctgcac ctaatgaatt
      661 tgatgtcatg tttaaactgg aagtccccag aattcaacta gaagaatatt ccaacactcg
      721 tgcatattac tttgtgaaat ttaaaagaaa tccgaaagaa aatcatctga gtcagttttt
      781 agaaggtgaa atattatcag cttctaagat gctgtcaaag tttaggaaaa tcattaagga
      841 agaaattaac gacattaaag atacagatgt catcatgaag aggaaaagag gagggagccc
      901 tgctgtaaca cttcttatta gtgaaaaaat atctgtggat ataaccctgg ctttggaatc
      961 aaaaagtagc tggcctgcta gcacccaaga aggcctgcgc attcaaaact ggctttcagc
     1021 aaaagttagg aagcaactac gactaaagcc attttacctt gtacccaagc atgcaaagga
     1081 aggaaatggt ttccaagaag aaacatggcg gctatccttc tctcacatcg aaaaggaaat
     1141 tttgaacaat catggaaaat ctaaaacgtg ctgtgaaaac aaagaagaga aatgttgcag
     1201 gaaagattgt ttaaaactaa tgaaatacct tttagaacag ctgaaagaaa ggtttaaaga
     1261 cgaagaacat ctggataaat tctcttctta tcatgtgaaa actgccttct ttcacgtatg
     1321 tacccagaac cctcaagaca gtcagtggga ccgcaaagac ctgggcctct gctttgataa
     1381 ctgcgtgaca tactttcttc agtgcctcag gacagaaaaa cttgagaatt attttattcc
     1441 tgaattcaat ctattctcta gcaacttaat tgacaaaaga agtaaagaat ttctgacaaa
     1501 gcaaattgaa tatgaaagaa acaatgagtt tccagttttt gatgaatttt aactcgagtc
     1561 tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc
     1621 agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1681 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
     1741 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
     1801 acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt ggcacttttc
     1861 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1921 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     1981 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2041 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     2101 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     2161 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     2221 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     2281 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa
     2341 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     2401 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     2461 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     2521 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     2581 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     2641 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     2701 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc
     2761 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2821 gaatgtattt agaaaaataa acaaataggc atgcagcgct cttccgcttc ctcgctcact
     2881 gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt cagctcactc aaaagcggta
     2941 atacggttat ccacagaatc aggggataaa gccggaaaga acatgtgagc aaaaagcaaa
     3001 gcaccggaag aagccaacgc cgcaggcgtt tttccatagg ctccgccccc ctgacgagca
     3061 tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg acaggactat aaagatacca
     3121 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3181 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgttg
     3241 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3301 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3361 cgacttatcg ccactggcag cagccattgg taactgattt agaggacttt gtcttgaagt
     3421 tatgcacctg ttaaggctaa actgaaagaa cagattttgg tgagtgcggt cctccaaccc
     3481 acttaccttg gttcaaagag ttggtagctc agcgaacctt gagaaaacca ccgttggtag
     3541 cggtggtttt tctttattta tgagatgatg aatcaatcgg tctatcaagt caacgaacag
     3601 ctattccgtt actctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     3661 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
     3721 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     3781 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3841 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     3901 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     3961 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     4021 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     4081 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     4141 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     4201 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     4261 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     4321 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     4381 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     4441 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     4501 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     4561 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     4621 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     4681 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     4741 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     4801 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     4861 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     4921 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     4981 tctcccttat gcgactcctg cattaggaaa ttaatacgac tcactata
//
