LOCUS       sequence-366530-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of mouse cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  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 7715)
  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-mouse cGAS:145-508aa-HA
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        323..526
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             527..702
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    765..964
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1165..1581
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1511..1533
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1549..1565
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1621..1970
                     /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    1629..1647
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1665..1683
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1701..1719
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1737..1755
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1773..1791
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1809..1827
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1845..1863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1909..1933
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3075..3101
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3135..3634
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3170..3193)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3567..3586
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3655..4254
                     /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(3655..3674)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4151..4171
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4435..4860
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4963..4985)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5121..5141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5126..5455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5306..5441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5477..5496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5577..5594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5748..6336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5828..5847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6507..7298,7299..7367))
                     /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     7130..7149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7368..7472)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7540..7558
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7658..7715,1..322)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
       61 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      121 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      181 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      241 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      301 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      361 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      421 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      481 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcaat aaaagagccc
      541 acaacccctc actcggcgcg ccagtcttcc gatagactgc gtcgcccggg tacccgtatt
      601 cccaataaag cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc ttgggagggt
      661 ctcctctgag tgattgacta cccacgacgg gggtctttca tttgggggct cgtccgggat
      721 ttggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact
      781 tatctgtgtc tgtccgattg tctagtgtct atgtttgatg ttatgcgcct gcgtctgtac
      841 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca
      901 cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc
      961 tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac
     1021 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc
     1081 cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg
     1141 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
     1201 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
     1261 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
     1321 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
     1381 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
     1441 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
     1501 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
     1561 ctcactcctt ctctaggcgc cggaattgaa gatctcgatg aggccctttc gtcttcactc
     1621 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
     1681 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
     1741 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
     1801 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
     1861 agaacgtatg tcgaggtagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
     1921 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatactt ggatccgcca
     1981 ccatgaagga accggacaag ctaaagaagg tgctggacaa attgagattg aaacgcaaag
     2041 atatctcgga ggcggccgag acggtgaata aagttgtgga acgcctgctg cgcagaatgc
     2101 agaaacggga gtcggagttc aaaggtgtgg agcagctgaa cactggcagc tactatgaac
     2161 atgtgaagat ttctgctcct aatgaatttg atgttatgtt taaactggaa gtccccagga
     2221 ttgagctaca agaatattat gaaacaggtg ctttctatct tgtgaaattc aaaagaattc
     2281 cacgaggaaa tccgctgagt catttcttag aaggggaagt attatcagct accaagatgc
     2341 tgtcaaagtt taggaaaatc attaaagaag aagttaaaga aatcaaagat atagatgtca
     2401 gtgtggagaa ggaaaaacca ggaagccctg ctgtaacact tcttatcagg aaccctgaag
     2461 aaatctctgt ggatataatt ctggctttgg agtcaaaagg cagctggcct attagtacca
     2521 aagaaggact acctattcaa ggctggctgg gcacaaaagt gaggaccaat ctaagacgag
     2581 agccgtttta tctcgtaccc aagaatgcaa aggatggaaa tagttttcaa ggagagacct
     2641 ggcgcctctc tttctctcac actgaaaagt acattttgaa taatcacggg atagagaaaa
     2701 catgctgtga atcttccgga gcaaaatgct gcagaaaaga atgtttaaaa ttaatgaaat
     2761 accttttgga acagttgaaa aaagagtttc aagagctgga tgcattctgt tcctaccatg
     2821 tgaaaactgc catctttcac atgtggaccc aggacccgca ggacagtcag tgggacccca
     2881 ggaacctcag ctcctgcttc gataagttgt tagcattctt tcttgagtgc ctcaggacag
     2941 agaaactgga tcattatttt attccaaagt tcaatctatt ctctcaagaa ctaattgacc
     3001 gaaaaagtaa agaatttcta tcgaagaaaa ttgaatatga aagaaataat gggtttccaa
     3061 tttttgacaa gctttaccca tacgatgttc cagattacgc ttgagcggcc gcgccggcgg
     3121 gcccgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     3181 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     3241 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     3301 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     3361 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     3421 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     3481 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     3541 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     3601 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc
     3661 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3721 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3781 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3841 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3901 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3961 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     4021 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4081 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     4141 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4201 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     4261 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc ggcgtctcga
     4321 gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga
     4381 gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     4441 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     4501 aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc
     4561 gaccctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4621 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4681 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     4741 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4801 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     4861 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
     4921 gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4981 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5041 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     5101 cggagtgtag atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5221 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5281 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5341 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5401 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5461 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5521 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     5581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6181 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     6841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     7141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7501 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7561 tttcgtcttc aagaattagc ttggccattg catacgttgt atccatatca taatatgtac
     7621 atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat
     7681 taatagtaat caattacggg gtcattagtt catag
//
