LOCUS       sequence-257766-        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"
     primer_bind     812..831
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(851..928)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     974..993
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        974..992
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    993..1017
                     /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             1073..1090
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1100..1117
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1130..1423
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             2555..2599
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2637..2655)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2651..2698
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2928..3743)
                     /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(2928..3743)
                     /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     3651..3670
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3744..3835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3851..4600)
                     /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(3861..3878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4101..4120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4796..5028,1..850))
                     /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 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
       61 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
      121 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
      181 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
      241 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
      301 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
      361 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
      421 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
      481 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
      541 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
      601 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
      661 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
      721 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
      781 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
      841 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
      901 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
      961 tgcattagga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     1021 gtagaaataa ttttgtttaa ctttaataag gagatatacc atgggcagca gccatcatca
     1081 tcatcatcac agcagcggcc tggtgccgcg cggcagccac atggcaagca tgtcggactc
     1141 agaagtcaat caagaagcta agccagaggt caagccagaa gtcaagcctg agactcacat
     1201 caatttaaag gtgtccgatg gatcttcaga gatcttcttc aagatcaaaa agaccactcc
     1261 tttaagaagg ctgatggaag cgttcgctaa aagacagggt aaggaaatgg actccttaag
     1321 attcttgtac gacggtatta gaatccaagc tgatcagacc cctgaagatt tggacatgga
     1381 ggataacgat attattgagg ctcacagaga acagattggt ggatccattc tcgtacggag
     1441 ggatgcggcg cctggggcct cgaagctccg ggcggttttg gagaagttga agctcagccg
     1501 cgatgatatc tccacggcgg cggggatggt gaaaggggtt gtggaccacc tgctgctcag
     1561 actgaagtgc gactccgcgt tcagaggcgt cgggctgctg aacaccggga gctactatga
     1621 gcacgtgaag atttctgcac ctaatgaatt tgatgtcatg tttaaactgg aagtccccag
     1681 aattcaacta gaagaatatt ccaacactcg tgcatattac tttgtgaaat ttaaaagaaa
     1741 tccgaaagaa aatcatctga gtcagttttt agaaggtgaa atattatcag cttctaagat
     1801 gctgtcaaag tttaggaaaa tcattaagga agaaattaac gacattaaag atacagatgt
     1861 catcatgaag aggaaaagag gagggagccc tgctgtaaca cttcttatta gtgaaaaaat
     1921 atctgtggat ataaccctgg ctttggaatc aaaaagtagc tggcctgcta gcacccaaga
     1981 aggcctgcgc attcaaaact ggctttcagc aaaagttagg aagcaactac gactaaagcc
     2041 attttacctt gtacccaagc atgcaaagga aggaaatggt ttccaagaag aaacatggcg
     2101 gctatccttc tctcacatcg aaaaggaaat tttgaacaat catggaaaat ctaaaacgtg
     2161 ctgtgaaaac aaagaagaga aatgttgcag gaaagattgt ttaaaactaa tgaaatacct
     2221 tttagaacag ctgaaagaaa ggtttaaaga cgaagaacat ctggataaat tctcttctta
     2281 tcatgtgaaa actgccttct ttcacgtatg tacccagaac cctcaagaca gtcagtggga
     2341 ccgcaaagac ctgggcctct gctttgataa ctgcgtgaca tactttcttc agtgcctcag
     2401 gacagaaaaa cttgagaatt attttattcc tgaattcaat ctattctcta gcaacttaat
     2461 tgacaaaaga agtaaagaat ttctgacaaa gcaaattgaa tatgaaagaa acaatgagtt
     2521 tccagttttt gatgaatttt aactcgagtc tggtaaagaa accgctgctg cgaaatttga
     2581 acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg ctgccaccgc
     2641 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2701 gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt
     2761 aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg
     2821 acgaccgggt ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2881 tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca
     2941 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3001 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3061 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     3121 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     3181 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     3241 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     3301 cgaaatacgc ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     3361 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3421 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3481 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     3541 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3601 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3661 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac
     3721 gtttcccgtt gaatatggct catactcttc ctttttcaat attattgaag catttatcag
     3781 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc
     3841 atgcagcgct cttccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     3901 cgagcggtgt cagctcactc aaaagcggta atacggttat ccacagaatc aggggataaa
     3961 gccggaaaga acatgtgagc aaaaagcaaa gcaccggaag aagccaacgc cgcaggcgtt
     4021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gccagaggtg
     4081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4201 cgtggcgctt tctcatagct cacgctgttg gtatctcagt tcggtgtagg tcgttcgctc
     4261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccattgg
     4381 taactgattt agaggacttt gtcttgaagt tatgcacctg ttaaggctaa actgaaagaa
     4441 cagattttgg tgagtgcggt cctccaaccc acttaccttg gttcaaagag ttggtagctc
     4501 agcgaacctt gagaaaacca ccgttggtag cggtggtttt tctttattta tgagatgatg
     4561 aatcaatcgg tctatcaagt caacgaacag ctattccgtt actctagatt tcagtgcaat
     4621 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     4681 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     4741 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4861 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     4921 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     4981 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcg
//
