LOCUS       sequence-366037-        7739 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 7739)
  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 7739)
  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-FLAG-human cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        325..528
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             529..704
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    767..966
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1167..1583
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1513..1535
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1551..1567
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1623..1972
                     /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    1631..1649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1667..1685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1703..1721
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1739..1757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1775..1793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1811..1829
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1847..1865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1911..1935
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1979..1988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1988..2011
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3101..3127
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3161..3660
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3196..3219)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3593..3612
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3681..4280
                     /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(3681..3700)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4177..4197
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4461..4886
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4989..5011)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5147..5167)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5152..5481
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5332..5467
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5503..5522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5603..5620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5774..6362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5854..5873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6533..7324,7325..7393))
                     /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     7156..7175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7394..7498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7566..7584
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7684..7739,1..324)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
       61 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      121 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      181 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      241 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      301 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      361 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      421 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      481 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc
      541 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
      601 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
      661 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
      721 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      781 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
      841 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      901 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      961 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     1021 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     1081 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     1141 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1201 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1261 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1321 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1381 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1441 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1501 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1561 ccctcactcc ttctctaggc gccggaattg aagatctcga tgaggccctt tcgtcttcac
     1621 tcgagtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat
     1681 agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc
     1741 cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc
     1801 tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat
     1861 agagaacgta tgtcgaggta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta
     1921 gtgaaccgtc agatcgcctg gagcaattcc acaacacttt tgtcttatac ttggatccgc
     1981 caccatggat tacaaggatg acgacgataa gcctggggcc tcgaagctcc gggcggtttt
     2041 ggagaagttg aagctcagcc gcgatgatat ctccacggcg gcggggatgg tgaaaggggt
     2101 tgtggaccac ctgctgctca gactgaagtg cgactccgcg ttcagaggcg tcgggctgct
     2161 gaacaccggg agctactatg agcacgtgaa gatttctgca cctaatgaat ttgatgtcat
     2221 gtttaaactg gaagtcccca gaattcaact agaagaatat tccaacactc gtgcatatta
     2281 ctttgtgaaa tttaaaagaa atccgaaaga aaatcctctg agtcagtttt tagaaggtga
     2341 aatattatca gcttctaaga tgctgtcaaa gtttaggaaa atcattaagg aagaaattaa
     2401 cgacattaaa gatacagatg tcatcatgaa gaggaaaaga ggagggagcc ctgctgtaac
     2461 acttcttatt agtgaaaaaa tatctgtgga tataaccctg gctttggaat caaaaagtag
     2521 ctggcctgct agcacccaag aaggcctgcg cattcaaaac tggctttcag caaaagttag
     2581 gaagcaacta cgactaaagc cattttacct tgtacccaag catgcaaagg aaggaaatgg
     2641 tttccaagaa gaaacatggc ggctatcctt ctctcacatc gaaaaggaaa ttttgaacaa
     2701 tcatggaaaa tctaaaacgt gctgtgaaaa caaagaagag aaatgttgca ggaaagattg
     2761 tttaaaacta atgaaatacc ttttagaaca gctgaaagaa aggtttaaag acaaaaaaca
     2821 tctggataaa ttctcttctt atcatgtgaa aactgccttc tttcacgtat gtacccagaa
     2881 ccctcaagac agtcagtggg accgcaaaga cctgggcctc tgctttgata actgcgtgac
     2941 atactttctt cagtgcctca ggacagaaaa acttgagaat tattttattc ctgaattcaa
     3001 tctattctct agcaacttaa ttgacaaaag aagtaaagaa tttctgacaa agcaaattga
     3061 atatgaaaga aacaatgagt ttccagtttt tgatgaattt tacccatacg atgttccaga
     3121 ttacgcttga gcggccgcgc cggcgggccc gaattctacc gggtagggga ggcgcttttc
     3181 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     3241 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     3301 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     3361 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     3421 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     3481 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     3541 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     3601 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     3661 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     3721 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     3781 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     3841 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     3901 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     3961 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     4021 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     4081 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     4141 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     4201 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     4261 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
     4321 cacgacccca tgcatcggcg tctcgagata tcagtggtcc aggctctagt tttgactcaa
     4381 caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt
     4441 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4501 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca
     4561 aggtcaggaa cagatggaac agggtcgacc ctagagaacc atcagatgtt tccagggtgc
     4621 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4681 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4741 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     4801 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     4861 ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc
     4921 agggaccacc gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt cggtgatgac
     4981 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5041 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     5101 gccatgaccc agtcacgtag cgatagcgga gtgtagatcc ggctgtggaa tgtgtgtcag
     5161 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5221 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5281 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5341 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5401 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5461 ggaggcctag gcttttgcaa aaagcttact ggcttaacta tgcggcatca gagcagattg
     5521 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5581 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6181 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6841 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7141 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7501 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     7561 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attagcttgg ccattgcata
     7621 cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat
     7681 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcat
//
