LOCUS       sequence-366578-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of human cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  AUTHORS   Mosallanejad K, Kennedy SN, Bahleda KM, Slavik KM, Zhou W, Govande 
            AA, Hancks DC, Kranzusch PJ, Kagan JC
  TITLE     Species-specific self-DNA detection mechanisms by mammalian cyclic 
            GMP-AMP synthases.
  JOURNAL   Sci Immunol. 2023 Jan 20;8(79):eabp9765. doi: 
            10.1126/sciimmunol.abp9765. Epub 2023 Jan 20.
  PUBMED    36662885
REFERENCE   2  (bases 1 to 8189)
  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: "Sci 
            Immunol."; date: "2023-01-20"; volume: "8(79):eabp9765. doi"; pages:
            " 10.1126/sciimmunol.abp9765. Epub 2023 Jan 20"
COMMENT     Sequence Label: pTRE3G-human cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1607..1956
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    1615..1633
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1651..1669
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1687..1705
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1723..1741
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1759..1777
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1795..1813
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1831..1849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1895..1919
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3535..3561
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3595..4094
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3630..3653)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4027..4046
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4115..4714
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4115..4134)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4611..4631
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4895..5320
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5423..5445)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5581..5601)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5586..5915
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5766..5901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5937..5956
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6037..6054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6208..6796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6288..6307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6967..7758,7759..7827))
                     /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     7590..7609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7828..7932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8000..8018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8118..8189,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tcgatgaggc cctttcgtct tcactcgagt ttactcccta
     1621 tcagtgatag agaacgtatg aagagtttac tccctatcag tgatagagaa cgtatgcaga
     1681 ctttactccc tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag
     1741 aacgtatgac cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta
     1801 tcagtgatag agaacgtata tccagtttac tccctatcag tgatagagaa cgtatgtcga
     1861 ggtaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac cgtcagatcg
     1921 cctggagcaa ttccacaaca cttttgtctt atacttggat ccgccaccat gcagccttgg
     1981 cacggaaagg ccatgcagag agcttccgag gccggagcca ctgcccccaa ggcttccgca
     2041 cggaatgcca ggggcgcccc gatggatccc accgagtctc cggctgcccc cgaggccgcc
     2101 ctgcctaagg cgggaaagtt cggccccgcc aggaagtcgg gatcccggca gaaaaagagc
     2161 gccccggaca cccaggagag gccgcccgtc cgcgcaactg gggcccgcgc caaaaaggcc
     2221 cctcagcgcg cccaggacac gcagccgtct gacgccacca gcgcccctgg ggcagagggg
     2281 ctggagcctc ctgcggctcg ggagccggct ctttccaggg ctggttcttg ccgccagagg
     2341 ggcgcgcgct gctccacgaa gccaagacct ccgcccgggc cctgggacgt gcccagcccc
     2401 ggcctgccgg tctcggcccc cattctcgta cggagggatg cggcgcctgg ggcctcgaag
     2461 ctccgggcgg ttttggagaa gttgaagctc agccgcgatg atatctccac ggcggcgggg
     2521 atggtgaaag gggttgtgga ccacctgctg ctcagactga agtgcgactc cgcgttcaga
     2581 ggcgtcgggc tgctgaacac cgggagctac tatgagcacg tgaagatttc tgcacctaat
     2641 gaatttgatg tcatgtttaa actggaagtc cccagaattc aactagaaga atattccaac
     2701 actcgtgcat attactttgt gaaatttaaa agaaatccga aagaaaatcc tctgagtcag
     2761 tttttagaag gtgaaatatt atcagcttct aagatgctgt caaagtttag gaaaatcatt
     2821 aaggaagaaa ttaacgacat taaagataca gatgtcatca tgaagaggaa aagaggaggg
     2881 agccctgctg taacacttct tattagtgaa aaaatatctg tggatataac cctggctttg
     2941 gaatcaaaaa gtagctggcc tgctagcacc caagaaggcc tgcgcattca aaactggctt
     3001 tcagcaaaag ttaggaagca actacgacta aagccatttt accttgtacc caagcatgca
     3061 aaggaaggaa atggtttcca agaagaaaca tggcggctat ccttctctca catcgaaaag
     3121 gaaattttga acaatcatgg aaaatctaaa acgtgctgtg aaaacaaaga agagaaatgt
     3181 tgcaggaaag attgtttaaa actaatgaaa taccttttag aacagctgaa agaaaggttt
     3241 aaagacaaaa aacatctgga taaattctct tcttatcatg tgaaaactgc cttctttcac
     3301 gtatgtaccc agaaccctca agacagtcag tgggaccgca aagacctggg cctctgcttt
     3361 gataactgcg tgacatactt tcttcagtgc ctcaggacag aaaaacttga gaattatttt
     3421 attcctgaat tcaatctatt ctctagcaac ttaattgaca aaagaagtaa agaatttctg
     3481 acaaagcaaa ttgaatatga aagaaacaat gagtttccag tttttgatga attttaccca
     3541 tacgatgttc cagattacgc ttgagcggcc gcgccggcgg gcccgaattc taccgggtag
     3601 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     3661 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     3721 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     3781 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     3841 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     3901 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     3961 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     4021 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     4081 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     4141 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4201 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4261 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4321 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4381 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4441 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4501 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4561 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4621 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4681 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     4741 gaccgaaagg agcgcacgac cccatgcatc ggcgtctcga gatatcagtg gtccaggctc
     4801 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     4861 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     4921 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     4981 agaagttcag atcaaggtca ggaacagatg gaacagggtc gaccctagag aaccatcaga
     5041 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     5101 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     5161 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     5221 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     5281 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     5341 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc tgcctcgcgc
     5401 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5461 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5521 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtag atccggctgt
     5581 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5641 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5761 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5821 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5881 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tactggctta actatgcggc
     5941 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6001 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6061 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6841 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7261 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7561 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     7621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7921 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7981 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattagc
     8041 ttggccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     8101 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     8161 gtcattagtt catagcccat atatggagt
//
