LOCUS       sequence-257764-        5686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of human cGAS protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5686)
  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 5686)
  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-cGAS
FEATURES             Location/Qualifiers
     source          1..5686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     384..403
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(423..500)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        546..564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    565..589
                     /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             645..662
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             672..689
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             702..995
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     promoter        2633..2651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2652..2676
                     /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             2785..2829
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2867..2885)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2881..2928
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3158..3973)
                     /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(3158..3973)
                     /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     3881..3900
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3974..4065)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4081..4830)
                     /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(4091..4108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4331..4350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5026..5686,1..422))
                     /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"
ORIGIN
        1 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
       61 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
      121 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
      181 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
      241 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
      301 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
      361 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
      421 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
      481 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
      541 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctgtagaaat
      601 aattttgttt aactttaata aggagatata ccatgggcag cagccatcat catcatcatc
      661 acagcagcgg cctggtgccg cgcggcagcc acatggcaag catgtcggac tcagaagtca
      721 atcaagaagc taagccagag gtcaagccag aagtcaagcc tgagactcac atcaatttaa
      781 aggtgtccga tggatcttca gagatcttct tcaagatcaa aaagaccact cctttaagaa
      841 ggctgatgga agcgttcgct aaaagacagg gtaaggaaat ggactcctta agattcttgt
      901 acgacggtat tagaatccaa gctgatcaga cccctgaaga tttggacatg gaggataacg
      961 atattattga ggctcacaga gaacagattg gtggatctat gcagccttgg cacggaaagg
     1021 ccatgcagag agcttccgag gccggagcca ctgcccccaa ggcttccgca cggaatgcca
     1081 ggggcgcccc gatggatccc aacgagtctc cggctgcccc cgaggccgcc ctgcctaagg
     1141 cgggaaagtt cggccccgcc aggaagtcgg gatcccggca gaaaaagagc gccccggaca
     1201 cccaggagag gccgcccgtc cgcgcaactg gggcccgcgc caaaaaggcc cctcagcgcg
     1261 cccaggacac gcagccgtct gacgccacca gcgcccctgg ggcagagggg ctggagcctc
     1321 ctgcggctcg ggagccggct ctttccaggg ctggttcttg ccgccagagg ggcgcgcgct
     1381 gctccacgaa gccaagaccc ccgcccgggc cctgggacgt gcccagcccc ggcctgccgg
     1441 tctcggcccc cattctcgta cggagggatg cggcgcctgg ggcctcgaag ctccgggcgg
     1501 ttttggagaa gttgaagctc agccgcgatg atatctccac ggcggcgggg atggtgaaag
     1561 gggttgtgga ccacctgctg ctcagactga agtgcgactc cgcgttcaga ggcgtcgggc
     1621 tgctgaacac cgggagctac tatgagcacg tgaagatttc tgcacctaat gaatttgatg
     1681 tcatgtttaa actggaagtc cccagaattc aactagaaga atattccaac actcgtgcat
     1741 attactttgt gaaatttaaa agaaatccga aagaaaatca tctgagtcag tttttagaag
     1801 gtgaaatatt atcagcttct aagatgctgt caaagtttag gaaaatcatt aaggaagaaa
     1861 ttaacgacat taaagataca gatgtcatca tgaagaggaa aagaggaggg agccctgctg
     1921 taacacttct tattagtgaa aaaatatctg tggatataac cctggctttg gaatcaaaaa
     1981 gtagctggcc tgctagcacc caagaaggcc tgcgcattca aaactggctt tcagcaaaag
     2041 ttaggaagca actacgacta aagccatttt accttgtacc caagcatgca aaggaaggaa
     2101 atggtttcca agaagaaaca tggcggctat ccttctctca catcgaaaag gaaattttga
     2161 acaatcatgg aaaatctaaa acgtgctgtg aaaacaaaga agagaaatgt tgcaggaaag
     2221 attgtttaaa actaatgaaa taccttttag aacagctgaa agaaaggttt aaagacaaaa
     2281 aacatctgga taaattctct tcttatcatg tgaaaactgc cttctttcac gtatgtaccc
     2341 agaaccctca agacagtcag tgggaccgca aagacctggg cctctgcttt gataactgcg
     2401 tgacatactt tcttcagtgc ctcaggacag aaaaacttga gaattatttt attcctgaat
     2461 tcaatctatt ctctagcaac ttaattgaca aaagaagtaa agaatttctg acaaagcaaa
     2521 ttgaatatga aagaaacaat gagtttccag tttttgatga attttgagcg gccgcataat
     2581 gcttaagtcg aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga
     2641 ctcactatag gggaattgtg agcggataac aattccccat cttagtatat tagttaagta
     2701 taagaaggag atatacatat ggcagatctc aattggatat cggccggcca cgcgatcgct
     2761 gacgtcggta ccctcgagtc tggtaaagaa accgctgctg cgaaatttga acgccagcac
     2821 atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc tgagcaataa
     2881 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     2941 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     3001 tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt
     3061 ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3121 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     3181 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3241 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     3301 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3361 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3421 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3481 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3541 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3601 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3661 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     3721 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     3781 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     3841 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     3901 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     3961 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc atgcagcgct
     4081 cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt
     4141 cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa gccggaaaga
     4201 acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt tttccatagg
     4261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg
     4321 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4381 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4441 tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4501 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4561 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg taactgattt
     4621 agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa cagattttgg
     4681 tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc agcgaacctt
     4741 gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg aatcaatcgg
     4801 tctatcaagt caacgaacag ctattccgtt actctagatt tcagtgcaat ttatctcttc
     4861 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     4921 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     4981 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5101 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5161 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5221 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5281 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     5341 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5401 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5461 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5521 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5581 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5641 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acatta
//
