LOCUS       sequence-366109-        7694 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 7694)
  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 7694)
  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 cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        334..537
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             538..713
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    776..975
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1176..1592
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1522..1544
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1560..1576
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1632..1981
                     /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    1640..1658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1676..1694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1712..1730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1748..1766
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1784..1802
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1820..1838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1856..1874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1920..1944
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1988..1997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3077..3103
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3137..3624
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3172..3195)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3557..3576
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3645..4244
                     /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(3645..3664)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4141..4161
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4425..4850
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4953..4975)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5111..5131)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5116..5445
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5296..5431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5467..5486
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5567..5584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5738..6326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5818..5837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6497..7288,7289..7357))
                     /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     7120..7139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7358..7462)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7530..7548
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7648..7694,1..333)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
       61 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      121 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      181 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      241 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      301 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      361 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      421 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      481 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcaa
      541 taaaagagcc cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg
      601 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc
      661 cttgggaggg tctcctctga gtgattgact acccacgacg ggggtctttc atttgggggc
      721 tcgtccggga tttggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
      781 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc
      841 tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
      901 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
      961 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
     1021 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
     1081 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
     1141 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
     1201 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1261 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1321 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1381 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1441 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1501 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1561 tttatccagc cctcactcct tctctaggcg ccggaattga agatctcgat gaggcccttt
     1621 cgtcttcact cgagtttact ccctatcagt gatagagaac gtatgaagag tttactccct
     1681 atcagtgata gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg
     1741 agtttactcc ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga
     1801 gaacgtatct acagtttact ccctatcagt gatagagaac gtatatccag tttactccct
     1861 atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta taaaagcaga
     1921 gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatact
     1981 tggatccgcc accatggaca agctaaagaa ggtgctggac aaattgagat tgaaacgcaa
     2041 agatatctcg gaggcggccg agacggtgaa taaagttgtg gaacgcctgc tgcgcagaat
     2101 gcagaaacgg gagtcggagt tcaaaggtgt ggagcagctg aacactggca gctactatga
     2161 acatgtgaag atttctgctc ctaatgaatt tgatgttatg tttaaactgg aagtccccag
     2221 gattgagcta caagaatatt atgaaacagg tgctttctat cttgtgaaat tcaaaagaat
     2281 tccacgagga aatccgctga gtcatttctt agaaggggaa gtattatcag ctaccaagat
     2341 gctgtcaaag tttaggaaaa tcattaaaga agaagttaaa gaaatcaaag atatagatgt
     2401 cagtgtggag aaggaaaaac caggaagccc tgctgtaaca cttcttatca ggaaccctga
     2461 agaaatctct gtggatataa ttctggcttt ggagtcaaaa ggcagctggc ctattagtac
     2521 caaagaagga ctacctattc aaggctggct gggcacaaaa gtgaggacca atctaagacg
     2581 agagccgttt tatctcgtac ccaagaatgc aaaggatgga aatagttttc aaggagagac
     2641 ctggcgcctc tctttctctc acactgaaaa gtacattttg aataatcacg ggatagagaa
     2701 aacatgctgt gaatcttccg gagcaaaatg ctgcagaaaa gaatgtttaa aattaatgaa
     2761 ataccttttg gaacagttga aaaaagagtt tcaagagctg gatgcattct gttcctacca
     2821 tgtgaaaact gccatctttc acatgtggac ccaggacccg caggacagtc agtgggaccc
     2881 caggaacctc agctcctgct tcgataagtt gttagcattc tttcttgagt gcctcaggac
     2941 agagaaactg gatcattatt ttattccaaa gttcaatcta ttctctcaag aactaattga
     3001 ccgaaaaagt aaagaatttc tatcgaagaa aattgaatat gaaagaaata atgggtttcc
     3061 aatttttgac aagctttacc catacgatgt tccagattac gcttgagcgg ccgcgccggc
     3121 gggcccgaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     3181 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     3241 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     3301 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     3361 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     3421 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     3481 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcggggcggg cgcccgaagg
     3541 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     3601 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     3661 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     3721 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     3781 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     3841 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     3901 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     3961 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     4021 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     4081 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4141 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4201 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc
     4261 cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc ggcgtctcga gatatcagtg
     4321 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     4381 tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     4441 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     4501 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc gaccctagag
     4561 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     4621 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     4681 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     4741 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     4801 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
     4861 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
     4921 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4981 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5041 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtag
     5101 atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5161 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5221 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5281 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5341 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5401 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tactggctta
     5461 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     5521 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     5581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6601 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6721 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6781 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     6841 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6901 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6961 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7021 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7081 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     7141 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7201 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7261 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7321 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     7501 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     7561 aagaattagc ttggccattg catacgttgt atccatatca taatatgtac atttatattg
     7621 gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat taatagtaat
     7681 caattacggg gtca
//
