LOCUS       sequence-366046-        7733 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 7733)
  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 7733)
  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:154-522aa-HA
FEATURES             Location/Qualifiers
     source          1..7733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             535..710
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    773..972
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1173..1589
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1519..1541
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1557..1573
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1629..1978
                     /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    1637..1655
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1673..1691
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1709..1727
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1745..1763
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1781..1799
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1817..1835
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1853..1871
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1917..1941
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1985..1994
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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..7733,1..330)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa
      541 aagagcccac aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta
      601 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt
      661 gggagggtct cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg
      721 tccgggattt ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
      781 cagcaactta tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc
      841 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
      901 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
      961 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     1021 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     1081 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     1141 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     1201 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     1261 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     1321 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1381 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1441 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     1501 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     1561 atccagccct cactccttct ctaggcgccg gaattgaaga tctcgatgag gccctttcgt
     1621 cttcactcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     1681 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     1741 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     1801 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     1861 agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct
     1921 cgtttagtga accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttgg
     1981 atccgccacc atggtacgga gggatgcggc 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 tta
//
