LOCUS       sequence-366040-        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-HA-human cGAS
FEATURES             Location/Qualifiers
     source          1..8189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        318..521
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             522..697
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    760..959
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1160..1576
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1506..1528
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1544..1560
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1616..1965
                     /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    1624..1642
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1660..1678
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1696..1714
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1732..1750
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1768..1786
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1804..1822
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1840..1858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1904..1928
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1981..2007
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3604..4103
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3639..3662)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4036..4055
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4124..4723
                     /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(4124..4143)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4620..4640
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4904..5329
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5432..5454)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5590..5610)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5595..5924
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5775..5910
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5946..5965
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6046..6063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6217..6805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6297..6316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6976..7767,7768..7836))
                     /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     7599..7618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7837..7941)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8009..8027
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8127..8189,1..317)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
       61 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      301 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      361 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      421 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      481 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcaataaaag agcccacaac
      541 ccctcactcg gcgcgccagt cttccgatag actgcgtcgc ccgggtaccc gtattcccaa
      601 taaagcctct tgctgtttgc atccgaatcg tggtctcgct gttccttggg agggtctcct
      661 ctgagtgatt gactacccac gacgggggtc tttcatttgg gggctcgtcc gggatttgga
      721 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct
      781 gtgtctgtcc gattgtctag tgtctatgtt tgatgttatg cgcctgcgtc tgtactagtt
      841 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg
      901 ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg
      961 aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa
     1021 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc
     1081 gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct
     1141 gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca
     1201 ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg
     1261 ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac
     1321 ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg
     1381 acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc
     1441 ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc
     1501 tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac
     1561 tccttctcta ggcgccggaa ttgaagatct cgatgaggcc ctttcgtctt cactcgagtt
     1621 tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt gatagagaac
     1681 gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta ctccctatca
     1741 gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt atctacagtt
     1801 tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt gatagagaac
     1861 gtatgtcgag gtaggcgtgt acggtgggcg cctataaaag cagagctcgt ttagtgaacc
     1921 gtcagatcgc ctggagcaat tccacaacac ttttgtctta tacttggatc cgccaccatg
     1981 tacccatacg atgttccaga ttacgctcag ccttggcacg gaaaggccat gcagagagct
     2041 tccgaggccg gagccactgc ccccaaggct tccgcacgga atgccagggg cgccccgatg
     2101 gatcccaccg agtctccggc tgcccccgag gccgccctgc ctaaggcggg aaagttcggc
     2161 cccgccagga agtcgggatc ccggcagaaa aagagcgccc cggacaccca ggagaggccg
     2221 cccgtccgcg caactggggc ccgcgccaaa aaggcccctc agcgcgccca ggacacgcag
     2281 ccgtctgacg ccaccagcgc ccctggggca gaggggctgg agcctcctgc ggctcgggag
     2341 ccggctcttt ccagggctgg ttcttgccgc cagaggggcg cgcgctgctc cacgaagcca
     2401 agacctccgc ccgggccctg ggacgtgccc agccccggcc tgccggtctc ggcccccatt
     2461 ctcgtacgga gggatgcggc gcctggggcc tcgaagctcc gggcggtttt ggagaagttg
     2521 aagctcagcc gcgatgatat ctccacggcg gcggggatgg tgaaaggggt tgtggaccac
     2581 ctgctgctca gactgaagtg cgactccgcg ttcagaggcg tcgggctgct gaacaccggg
     2641 agctactatg agcacgtgaa gatttctgca cctaatgaat ttgatgtcat gtttaaactg
     2701 gaagtcccca gaattcaact agaagaatat tccaacactc gtgcatatta ctttgtgaaa
     2761 tttaaaagaa atccgaaaga aaatcctctg agtcagtttt tagaaggtga aatattatca
     2821 gcttctaaga tgctgtcaaa gtttaggaaa atcattaagg aagaaattaa cgacattaaa
     2881 gatacagatg tcatcatgaa gaggaaaaga ggagggagcc ctgctgtaac acttcttatt
     2941 agtgaaaaaa tatctgtgga tataaccctg gctttggaat caaaaagtag ctggcctgct
     3001 agcacccaag aaggcctgcg cattcaaaac tggctttcag caaaagttag gaagcaacta
     3061 cgactaaagc cattttacct tgtacccaag catgcaaagg aaggaaatgg tttccaagaa
     3121 gaaacatggc ggctatcctt ctctcacatc gaaaaggaaa ttttgaacaa tcatggaaaa
     3181 tctaaaacgt gctgtgaaaa caaagaagag aaatgttgca ggaaagattg tttaaaacta
     3241 atgaaatacc ttttagaaca gctgaaagaa aggtttaaag acaaaaaaca tctggataaa
     3301 ttctcttctt atcatgtgaa aactgccttc tttcacgtat gtacccagaa ccctcaagac
     3361 agtcagtggg accgcaaaga cctgggcctc tgctttgata actgcgtgac atactttctt
     3421 cagtgcctca ggacagaaaa acttgagaat tattttattc ctgaattcaa tctattctct
     3481 agcaacttaa ttgacaaaag aagtaaagaa tttctgacaa agcaaattga atatgaaaga
     3541 aacaatgagt ttccagtttt tgatgaattt tgagcggccg cgccggcggg cccgaattct
     3601 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     3661 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     3721 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     3781 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     3841 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     3901 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     3961 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     4021 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     4081 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc
     4141 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4201 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4261 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4321 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4381 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4441 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4501 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4561 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4621 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4681 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     4741 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gcgtctcgag atatcagtgg
     4801 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     4861 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4921 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     4981 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagggtcg accctagaga
     5041 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     5101 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     5161 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     5221 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     5281 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     5341 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggct
     5401 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5461 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5521 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtaga
     5581 tccggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5641 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5701 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5761 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5821 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5881 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt actggcttaa
     5941 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6001 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     6061 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6121 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6181 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6241 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6301 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6361 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6421 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6481 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6541 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6601 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6661 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6721 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6781 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6961 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7021 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7081 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7141 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7201 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7261 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     7321 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7381 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7441 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7501 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7561 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     7621 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7681 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7741 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7801 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7861 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7921 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7981 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     8041 agaattagct tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
     8101 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     8161 aattacgggg tcattagttc atagcccat
//
