LOCUS       sequence-366054-        7709 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 7709)
  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 7709)
  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-human cGAS:162-522aa-HA
FEATURES             Location/Qualifiers
     source          1..7709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        315..518
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             519..694
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    757..956
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1157..1573
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1503..1525
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1541..1557
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1613..1962
                     /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    1621..1639
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1657..1675
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1693..1711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1729..1747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1765..1783
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1801..1819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1837..1855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1901..1925
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1969..1978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3061..3087
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3121..3620
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3156..3179)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3553..3572
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3641..4240
                     /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(3641..3660)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4137..4157
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4421..4846
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4949..4971)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5107..5127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5112..5441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5292..5427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5463..5482
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5563..5580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5734..6322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5814..5833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6493..7284,7285..7353))
                     /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     7116..7135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7354..7458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7526..7544
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7644..7709,1..314)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
       61 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      121 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      181 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      241 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      301 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      361 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      421 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      481 gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc ccacaacccc
      541 tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta ttcccaataa
      601 agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg gtctcctctg
      661 agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg atttggagac
      721 ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg
      781 tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt actagttagc
      841 taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg
      901 caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag
      961 ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag acgagaacct
     1021 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc
     1081 ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta
     1141 tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct taggtcactg
     1201 gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt
     1261 taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt
     1321 taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca
     1381 cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg
     1441 ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct
     1501 cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc
     1561 ttctctaggc gccggaattg aagatctcga tgaggccctt tcgtcttcac tcgagtttac
     1621 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
     1681 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
     1741 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
     1801 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
     1861 tgtcgaggta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc
     1921 agatcgcctg gagcaattcc acaacacttt tgtcttatac ttggatccgc caccatggcc
     1981 tcgaagctcc gggcggtttt ggagaagttg aagctcagcc gcgatgatat ctccacggcg
     2041 gcggggattg tgaaaggggt tgtggaccac ctgctgctca gactgaagtg cgactccgcg
     2101 ttcagaggcg tcgggctgct gaacaccggg agctactatg agcacgtgaa gatttctgca
     2161 cctaatgaat ttgatgtcat gtttaaactg gaagtcccca gaattcaact agaagaatat
     2221 tccaacactc gtgcatatta ctttgtgaaa tttaaaagaa atccgaaaga aaatcctctg
     2281 agtcagtttt tagaaggtga aatattatca gcttctaaga tgctgtcaaa gtttaggaaa
     2341 atcattaagg aagaaattaa cgacattaaa gatacagatg tcatcatgaa gaggaaaaga
     2401 ggagggagcc ctgctgtaac acttcttatt agtgaaaaaa tatctgtgga tataaccctg
     2461 gctttggaat caaaaagtag ctggcctgct agcacccaag aaggcctgcg cattcaaaac
     2521 tggctttcag caaaagttag gaagcaacta cgactaaagc cattttacct tgtacccaag
     2581 catgcaaagg aaggaaatgg tttccaagaa gaaacatggc ggctatcctt ctctcacatc
     2641 gaaaaggaaa ttttgaacaa tcatggaaaa tctaaaacgt gctgtgaaaa caaagaagag
     2701 aaatgttgca ggaaagattg tttaaaacta atgaaatacc ttttagaaca gctgaaagaa
     2761 aggtttaaag acaaaaaaca tctggataaa ttctcttctt atcatgtgaa aactgccttc
     2821 tttcacgtat gtacccagaa ccctcaagac agtcagtggg accgcaaaga cctgggcctc
     2881 tgctttgata actgcgtgac atactttctt cagtgcctca ggacagaaaa acttgagaat
     2941 tattttattc ctgaattcaa tctattctct agcaacttaa ttgacaaaag aagtaaagaa
     3001 tttctgacaa agcaaattga atatgaaaga aacaatgagt ttccagtttt tgatgaattt
     3061 tacccatacg atgttccaga ttacgcttga gcggccgcgc cggcgggccc gaattctacc
     3121 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     3181 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     3241 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     3301 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     3361 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     3421 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     3481 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     3541 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     3601 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac
     3661 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     3721 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     3781 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     3841 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     3901 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     3961 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4021 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4081 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4141 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4201 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     4261 gcgcccgacc gaaaggagcg cacgacccca tgcatcggcg tctcgagata tcagtggtcc
     4321 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     4381 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4441 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga
     4501 gaatagagaa gttcagatca aggtcaggaa cagatggaac agggtcgacc ctagagaacc
     4561 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     4621 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     4681 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     4741 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     4801 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg
     4861 ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa gctggctgcc
     4921 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4981 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5041 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtagatcc
     5101 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5161 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5221 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5281 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5341 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5401 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttact ggcttaacta
     5461 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5521 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     5581 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6781 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     6841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7081 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     7141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7501 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     7561 attagcttgg ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc
     7621 atgtccaaca ttaccgccat gttgacattg attattgact agttattaat agtaatcaat
     7681 tacggggtca ttagttcata gcccatata
//
