LOCUS       sequence-366078-        8174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of orangutan cGAS gene in mammalian
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8174)
  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 8174)
  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-orangutan cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        330..533
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             534..709
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    772..971
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1172..1588
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1518..1540
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1556..1572
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1628..1977
                     /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    1636..1654
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1672..1690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1708..1726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1744..1762
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1780..1798
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1816..1834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1852..1870
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1916..1940
                     /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..8174,1..329)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
       61 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      121 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      181 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      241 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      301 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      361 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      421 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      481 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcaataaa
      541 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      601 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      661 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      721 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      781 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      841 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      901 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      961 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     1021 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     1081 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1141 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1201 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1261 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1321 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1381 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1441 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1501 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1561 tccagccctc actccttctc taggcgccgg aattgaagat ctcgatgagg ccctttcgtc
     1621 ttcactcgag tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca
     1681 gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt
     1741 tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac
     1801 gtatctacag tttactccct atcagtgata gagaacgtat atccagttta ctccctatca
     1861 gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg cgcctataaa agcagagctc
     1921 gtttagtgaa ccgtcagatc gcctggagca attccacaac acttttgtct tatacttgga
     1981 tccgccacca tgcagccttg gcacggaaag gccatgcgga gagcttccga ggccggagcc
     2041 actgcccccg aggcttccgc gcggagtgcc aggggcgccc cgacggatcc caacgagtct
     2101 ccggctgtcc ccgaggccgc cctgcctaag gcgggaaagt tcggccccgc caggaagtca
     2161 ggatcccggc agaaaaagag cggcccggac acccaggaga ggccgcccgg ccgcgcaact
     2221 ggggcccgcg ccaaaaaggc ccctcagcgc gcccaggaca cgcagccctc tgacgccacc
     2281 agcgcccctg gggcagaggg gctggagcca actgcggctc gggagccggc tctttccagg
     2341 gctggttctt gccgccaaag gggcgcgcgc tgctctccga agccaagacc cccgccccgg
     2401 ccctgggacg tgcccagtct cggcctgccg gtctcggccc ccatatggag ggatgcggcg
     2461 cctggggcct cgaagctccg ggcggttttg gagaagttga agctcagccg cctggaaatc
     2521 tccaaggcgg cggaggtggt gaacagggtt gtggaccacc tgctgcgcag actgcagaag
     2581 tacaactccg agttcagagg cgtcgggctg ctgcgcaccg ggagctacta tgagcacgtg
     2641 aagatttctg cacctaatga atttgatgtc atgtttaaac tggaagtccc cagaattcaa
     2701 ctagaagaat attccaacac tggtgcatat tactttgtga aatttaaaag aaatccgaaa
     2761 gaaaatcctc tgagtcagtt tttagaaggt gaaatattat cagcttctaa gatgctgtca
     2821 aagtttagga aaatcattaa ggaagaaatt aacaacatta aagatacaga tgtcatcatg
     2881 aagaggaaaa gaggagggag ccctgctgta acacttctta ttagtgaaga aatatctgtg
     2941 gatataaccc tggctttgga atcaaaaagt agctggcctg ctagcaccca agaaggcctg
     3001 cccattaaaa actggctttc agcaaaagtt aggaagcaac taagactaaa gccattttac
     3061 cttgtaccca agcatgcaaa ggaaggaaat ggtttccaag aagaaacatg gcggctatcc
     3121 ttctctcaca tcgaaaagga aattttgaac aatcatggaa aatctaaaac gtgctgtgaa
     3181 gacaaagaag tgaaatgttg caggaaagat tgtttaaaac taatgaaata ccttttagaa
     3241 cagctgaaag aaaggtttaa agacaaaaaa catctggata aattctcttc ttatcatgtg
     3301 aaaactgcct tctttcacgt atgtacccag aaccctcaag acagtcagtg ggaccccaaa
     3361 gacctgggcc tctgctttga taactgcgtg acatactttc ttcagtgcct caggacagaa
     3421 caacttggga attattttat tcctggattc aatctattct ctagcaactt 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 ttag
//
