LOCUS       sequence-366073-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of rhesus macaque 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-rhesus macaque cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        324..527
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             528..703
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    766..965
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1166..1582
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1512..1534
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1550..1566
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1622..1971
                     /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    1630..1648
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1666..1684
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1702..1720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1738..1756
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1774..1792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1810..1828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1846..1864
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1910..1934
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3550..3576
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3610..4109
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3645..3668)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4042..4061
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4130..4729
                     /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(4130..4149)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4626..4646
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4910..5335
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5438..5460)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5596..5616)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5601..5930
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5781..5916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5952..5971
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6052..6069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6223..6811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6303..6322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6982..7773,7774..7842))
                     /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     7605..7624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7843..7947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8015..8033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8133..8189,1..323)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
       61 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      361 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      421 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      481 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcaa taaaagagcc
      541 cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg gtacccgtat
      601 tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg
      661 tctcctctga gtgattgact acccacgacg ggggtctttc atttgggggc tcgtccggga
      721 tttggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac
      781 ttatctgtgt ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc tgcgtctgta
      841 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
      901 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
      961 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     1021 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     1081 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
     1141 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     1201 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     1261 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     1321 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     1381 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     1441 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     1501 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     1561 cctcactcct tctctaggcg ccggaattga agatctcgat gaggcccttt cgtcttcact
     1621 cgagtttact ccctatcagt gatagagaac gtatgaagag tttactccct atcagtgata
     1681 gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg agtttactcc
     1741 ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga gaacgtatct
     1801 acagtttact ccctatcagt gatagagaac gtatatccag tttactccct atcagtgata
     1861 gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag
     1921 tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatact tggatccgcc
     1981 accatgcagc ctcggcaagg aaaggccacg cggagagctt ccgaggccgg agccactgtc
     2041 cccgaggctt ccgcgcggag tgccaggggg gcccggaccg accgcaacga gtctccggct
     2101 gcccccgaag ccgccctgcc cgaggcggga aagttcggcc ccgccaggaa gccgggatcc
     2161 gggcagaaga agagcccccc ggacacccag gagcggtcgc ccgtccgcgc aactggggcc
     2221 cgcgctaaaa aggcccctca gcgcgcccag gacgcgcggc cctctgactc caccagcgcc
     2281 gctggggcgg aggggctgga gcctcctgcg gctcgggaac cggctgtttc cagggctggt
     2341 tctcgccgcc agaggggcgc gcgcggctct ccgaagccaa gacccctgcc cgggccccgg
     2401 gacgtgccca accccggcct gccggtctcg gtccccattc tggtacagag ggatgcggcg
     2461 cctggggcca ggaagctccg ggcggttttg gagaagttaa ggctcagcca ccaggacatc
     2521 tccaaggcgg cgaaggtggt gaatggggtt gtgggccacc tgctgcgcag attgaagtgc
     2581 gagcccgagt tcggaggcgt cgagcagctg cacaccggga gctactacga gcacgtgaag
     2641 atttctgcac ctaatgaatt cgatgtcatg tttaaactgc aagtccccag aattcagcta
     2701 gaagaatatt ctgacactgg tgcatattac tttgtgaaat ttaaaagaaa tccagaagga
     2761 aatcctctga gcctgttttt agaagatgaa atattatcag cttctaagat gctgtcaaag
     2821 tttaggaaaa tcattaagga agaaattaac aacattaaag atgcagatgt catcatgaag
     2881 aggaaaagac cagggagccc tgctgtaaca cttcttatta ataaagaaat atctgtggat
     2941 ataaccctgg ctttggaatt aaaaactagc tggcctgcca gcacccaaga aggcctggcc
     3001 attaaaaact ggctttcagc aaaagttagg cagcaactaa gactaaagcc gttttacctt
     3061 gtacccaagc atgcaaagga aggaaatggt ttccaagagg aaacatggcg gctatccttc
     3121 tctcacatcg aaaaggaaat tttgagcaat catggaaaat ctaaaacgtg ctgtgaaaac
     3181 aaagaaatga aatgttgcag gaaagattgt ttaaaactaa tgaaatacct tttagaacag
     3241 ctgaaagaaa agtttaaaga caaaaaacat ctggataaat tctcttctta tcatgtgaaa
     3301 actgccttct ttcacctatg taccgagaac cctcaagaca gtcagtggga ccccaaagac
     3361 ctgggcctct gctttgataa ctgcgtgaca tactttcttg agtgcctcag gacagaacaa
     3421 cttaagaatt attttattcc tggattcaat ctattctcta gcaacttaat tgacaaaaca
     3481 agtaaagaat ttctgtcaaa gcaaattgaa tatgaaagaa acaaggagtt tccagttttt
     3541 ggtgaatttt acccatacga tgttccagat tacgcttgag cggccgcgcc ggcgggcccg
     3601 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     3661 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     3721 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     3781 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     3841 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     3901 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     3961 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     4021 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     4081 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta
     4141 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4201 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4261 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4321 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4381 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4441 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4501 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4561 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4621 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4681 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     4741 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcggcgt ctcgagatat
     4801 cagtggtcca ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg
     4861 agccatagat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4921 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     4981 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gggtcgaccc
     5041 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     5101 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     5161 aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc
     5221 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     5281 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatttgggg
     5341 gctcgtccgg gatcgggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
     5401 ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     5461 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     5521 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     5581 tgtagatccg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5761 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5821 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     5881 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttactg
     5941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     6001 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     6061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6661 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7261 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     7321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     7621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     7981 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     8041 tcttcaagaa ttagcttggc cattgcatac gttgtatcca tatcataata tgtacattta
     8101 tattggctca tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata
     8161 gtaatcaatt acggggtcat tagttcata
//
