LOCUS       sequence-366072-        8264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of marmoset cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8264)
  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 8264)
  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-marmoset cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        311..514
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             515..690
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    753..952
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1153..1569
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1499..1521
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1537..1553
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1609..1958
                     /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    1617..1635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1653..1671
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1689..1707
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1725..1743
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1761..1779
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1797..1815
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1833..1851
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1897..1921
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3612..3638
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3672..4171
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3707..3730)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4104..4123
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4192..4791
                     /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(4192..4211)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4688..4708
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4972..5397
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5500..5522)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5658..5678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5663..5992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5843..5978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6014..6033
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6114..6131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6285..6873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6365..6384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7044..7835,7836..7904))
                     /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     7667..7686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7905..8009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8077..8095
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8195..8264,1..310)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
       61 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      121 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      181 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      241 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      301 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      361 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      421 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      481 gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac aacccctcac
      541 tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc caataaagcc
      601 tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct cctctgagtg
      661 attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt ggagacccct
      721 gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta tctgtgtctg
      781 tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta gttagctaac
      841 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
      901 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
      961 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     1021 cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt
     1081 ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     1141 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     1201 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     1261 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     1321 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     1381 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     1441 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     1501 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     1561 ctaggcgccg gaattgaaga tctcgatgag gccctttcgt cttcactcga gtttactccc
     1621 tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag aacgtatgca
     1681 gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta tcagtgatag
     1741 agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca gtttactccc
     1801 tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag aacgtatgtc
     1861 gaggtaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga accgtcagat
     1921 cgcctggagc aattccacaa cacttttgtc ttatacttgg atccgccacc atgcagcctc
     1981 gacgcggaaa ggccacgcgg agagcttccc aggcccgagc caccgctccc gaggcttctg
     2041 cgccaagcgc caggggcgcc cggacggacc caaaccagtc tctggctgct ccggaggccg
     2101 cccggcccca cgcgggaaag ttcggccccg ccagggggtc gggtagtagg cagaagaaga
     2161 acgcccagga cacccaggaa aggccgccag cccgtgagct gggggcccgc cccaagaagg
     2221 tccctcagcg cgccgaggac gcgcggctct cggacgccat cagcgcctct ggggcggagg
     2281 cgggagagcc tcccgaggct cccgagccgg ctttttccag agctggttct cgccgcccga
     2341 cgggtgcgcg ctgctctgcg gagcccagac ccccgcccgg gccccgggac ttgcccagcc
     2401 ccggcctgcc ggtctccggc cccatcctcg gccgaaggga gtcggcgccg cggaccagga
     2461 agctccgggc ggttttagag aagttaaggc taagtcgcga cgatgtctcc aacgcggcca
     2521 aggtggtgaa cagcgttgtg gaccacctgc tgggcagact gcagcagtgc gagtccgagt
     2581 tcaaaggcaa gttcaaagtg gagtcgctgc acaccgggag ctactatgag cgcgtgaaga
     2641 tttctacacc taatgaattt gatgtcatgt ttaaactgga actccccaga atccaactag
     2701 aagaatattc caacactggt gcatattact ttgtgaaatt caaaagaaat ctgaaagaaa
     2761 atcctctgag acaattttta gaagatgaaa tattatcagc ttctaagatg ctgtctaagt
     2821 ttaggaaaat cattaaggaa gaaattaaaa acatgcaaga tacagatgtc atcatggaga
     2881 ggaaaagaag agggagccct gctgtaacac ttcttattag aaaagaaata tctgtggata
     2941 taatcctggc tttggaatca aaaagtagct ggcctgctag cacccagaaa ggcctgccca
     3001 tcaaaaactg gctttcagcg aaagttagga cgaagctaag acgagagcca ttttaccttg
     3061 tacccaagca tgcaaaggaa ggaaatagtt tccaagaaga aacatggcgg ctatccttct
     3121 ctcacattga aaaggaaatt ttgagggatc atggacaatc taaaacgtgt tgtgaagaga
     3181 aaggagtgaa atgttgcagg aaagattgtt taaaactgat gaaatacctt ttagaacggc
     3241 tgaaagaaaa gtttaaaagt ggaaaacata tggataaatt ctgttcttat catgtgaaaa
     3301 ctgctttctt tcacgtatgt accgagaacc ctcaagacag tcagtgggac cccaaagatc
     3361 tggagctctg ctttgataac tgcctgacat actttcttca gtgcctcagg acagaaaaac
     3421 ttgaacatta tttcattcct gcattcaatc tattctctag caacttaatt gacaaaaaaa
     3481 gtaaagaatt tctgtcaaag caaattgaac atgaaagaaa caatgagttt ccagtttttg
     3541 gtgaatttcg agattctatg tttagaaaaa atttgagtga ctctcagtcc tggagaatac
     3601 caaaaaaatt ttacccatac gatgttccag attacgcttg agcggccgcg ccggcgggcc
     3661 cgaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     3721 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     3781 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     3841 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     3901 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     3961 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     4021 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc
     4081 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     4141 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag
     4201 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4261 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4321 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4381 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4441 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4501 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4561 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4621 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4681 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4741 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     4801 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcggc gtctcgagat
     4861 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta
     4921 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     4981 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     5041 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagggtcgac
     5101 cctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     5161 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     5221 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     5281 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     5341 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     5401 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     5461 agctggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5521 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5581 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     5641 agtgtagatc cggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     5701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     5761 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5821 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5881 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5941 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     6001 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     6061 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     6121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6961 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7021 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7081 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7141 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7201 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7261 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7321 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     7381 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7441 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7501 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7561 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7621 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     7681 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7741 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7801 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7861 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7921 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7981 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8041 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8101 cgtcttcaag aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt
     8161 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
     8221 tagtaatcaa ttacggggtc attagttcat agcccatata tgga
//
