LOCUS       sequence-366438-        8171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of chimpanzee cGAS gene in 
            mammalian cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8171)
  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 8171)
  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-chimpanzee cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        333..536
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             537..712
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    775..974
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1175..1591
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1521..1543
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1559..1575
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1631..1980
                     /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    1639..1657
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1675..1693
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1711..1729
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1747..1765
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1783..1801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1819..1837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1855..1873
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1919..1943
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3553..3579
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3613..4100
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3648..3671)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4033..4052
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4121..4720
                     /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(4121..4140)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4617..4637
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4901..5326
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5429..5451)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5587..5607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5592..5921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5772..5907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5943..5962
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6043..6060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6214..6802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6294..6313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6973..7764,7765..7833))
                     /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     7596..7615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7834..7938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8006..8024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8124..8171,1..332)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
       61 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      121 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      181 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      241 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      301 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      361 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      421 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      481 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcaat
      541 aaaagagccc acaacccctc actcggcgcg ccagtcttcc gatagactgc gtcgcccggg
      601 tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc
      661 ttgggagggt ctcctctgag tgattgacta cccacgacgg gggtctttca tttgggggct
      721 cgtccgggat ttggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
      781 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgtttgatg ttatgcgcct
      841 gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      901 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      961 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
     1021 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
     1081 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
     1141 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
     1201 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1261 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1321 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1381 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1441 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1501 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1561 ttatccagcc ctcactcctt ctctaggcgc cggaattgaa gatctcgatg aggccctttc
     1621 gtcttcactc gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta
     1681 tcagtgatag agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga
     1741 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
     1801 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
     1861 tcagtgatag agaacgtatg tcgaggtagg cgtgtacggt gggcgcctat aaaagcagag
     1921 ctcgtttagt gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatactt
     1981 ggatccgcca ccatgcagcc ttggcacgga aaggccatgc agagagcttc cgaggccgga
     2041 gccactgccc ccaaggcttc cgcgcggaat gccaggggcg ccacgacgta ccccaacgag
     2101 tctccggctg cccccgaggc cgccctgcct aaggcgggaa agttcggccc cgccaggaag
     2161 tcgggtagtc ggcagaaaaa gagcgccccg gacacccagg agaggccgcc cgtccgcgca
     2221 accggggccc gcgccaaaaa ggcccctcag cgcgcccagg acacgcagcc gtctgacgcc
     2281 accagcgccc ctggggcaga ggggctggag cctcctgcgg ctcgggaccc ggctctttcc
     2341 agggctggtt cttgccgcca gaggggcgcg cgctgctcta cgaagccaag acccccgccc
     2401 gggccctggg acgtgcccag ccccggcctg ccggtctcgg cccccattct cgtaaggagg
     2461 gatgcggcgc ctggggcctc gaagctccgg gcggttttgg agaagttgaa gctcagccgc
     2521 gatgatatct ccacggcggc ggggatggtg aaaggggttg tggaccacct gctgctcaga
     2581 ctgaagtgcg actccgcgtt cagaggcgtc gggctgctga acaccgggag ctactatgag
     2641 cacgtgaaga tttctgcacc taatgaattt gatgtcatgt ttaaactgga agtccccaga
     2701 atccaactag aagaatattc caacactcgt gcatattact ttgtgaaatt taaaagaaat
     2761 ccgaaagaaa atcctctgag tcagttttta gaaggtgaaa tattatcagc ttctaagatg
     2821 ctgtcaaagt ttaggaaaat cattaaggaa gaaattaaca acattaaaga tgtcatcatg
     2881 aagaggaaaa gaggagggag ccctgctgta acacttctta ttagtgaaaa aatatctgtg
     2941 gatataaccc tggctttgga atcaaaaagt agctggcctg ctagcacaca agaaggcctg
     3001 cccattaaaa actggctttc agcaaaagtt aggaagcaac taagactaaa gccattttac
     3061 cttgtaccca agcatgcaaa ggaaggaaat ggtttccaag aagaaacatg gcggctatcc
     3121 ttctctcaca tcgaaaagga aattttgaac aatcatggaa aatctaaaac gtgctgtgaa
     3181 atcaaagaag cgaaatgttg caggaaagat tgtttaaaac taatgaaata cattttagaa
     3241 cagctgaaag aaaagtttaa agacaaaaaa catctggata aattctcttc ttatcatgtg
     3301 aaaactgcct tctttcacgt atgtaccgag aaccctcaag acagtcagtg ggaccgcaaa
     3361 gacctgggcc tctgctttga taactgcgtg acatactttc ttcagtgcct caggatagaa
     3421 aaacttgaga attattttat tcctgaattt aatctattct ctagcgactt aattgacaaa
     3481 agaagtaaag aatttctgac aaagcaaatt gaatatgaaa gaaacaatga gtttccagtt
     3541 tttgatgaat tttacccata cgatgttcca gattacgctt gagcggccgc gccggcgggc
     3601 ccgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     3661 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     3721 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     3781 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     3841 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     3901 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     3961 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     4021 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     4081 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac
     4141 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4201 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4261 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4321 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4381 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4441 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4501 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4561 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4621 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4681 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     4741 gcgcccgacc gaaaggagcg cacgacccca tgcatcggcg tctcgagata tcagtggtcc
     4801 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     4861 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4921 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     4981 gaatagagaa gttcagatca aggtcaggaa cagatggaac agggtcgacc ctagagaacc
     5041 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     5101 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     5161 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     5221 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     5281 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg
     5341 ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa gctggctgcc
     5401 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5461 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5521 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtagatcc
     5581 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5641 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5761 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5821 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5881 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttact ggcttaacta
     5941 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6001 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     6061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7261 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     7321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7561 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     7621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7981 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     8041 attagcttgg ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc
     8101 atgtccaaca ttaccgccat gttgacattg attattgact agttattaat agtaatcaat
     8161 tacggggtca t
//
