LOCUS       sequence-366050-        7721 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 7721)
  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 7721)
  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:158-522aa-HA
FEATURES             Location/Qualifiers
     source          1..7721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        300..503
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             504..679
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    742..941
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1142..1558
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1488..1510
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1526..1542
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1598..1947
                     /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    1606..1624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1642..1660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1678..1696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1714..1732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1750..1768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1786..1804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1822..1840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1886..1910
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1954..1963
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1958..1967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3058..3084
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3118..3617
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3153..3176)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3550..3569
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3638..4237
                     /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(3638..3657)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4134..4154
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4418..4843
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4946..4968)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5104..5124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5109..5438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5289..5424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5460..5479
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5560..5577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5731..6319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5811..5830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6490..7281,7282..7350))
                     /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     7113..7132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7351..7455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7523..7541
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7641..7721,1..299)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
       61 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      121 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      181 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      241 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      301 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      361 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      421 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      481 gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact cggcgcgcca
      541 gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt
      601 gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
      661 acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg cccagggacc
      721 accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct
      781 agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact agctctgtat
      841 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
      901 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
      961 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     1021 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     1081 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     1141 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     1201 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     1261 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     1321 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     1381 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     1441 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     1501 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     1561 aattgaagat ctcgatgagg ccctttcgtc ttcactcgag tttactccct atcagtgata
     1621 gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc
     1681 ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga
     1741 ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata
     1801 gagaacgtat atccagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt
     1861 gtacggtggg cgcctataaa agcagagctc gtttagtgaa ccgtcagatc gcctggagca
     1921 attccacaac acttttgtct tatacttgga tccgccacca tggcggcgcc tggggcctcg
     1981 aagctccggg cggttttgga gaagttgaag ctcagccgcg atgatatctc cacggcggcg
     2041 gggatggtga aaggggttgt ggaccacctg ctgctcagac tgaagtgcga ctccgcgttc
     2101 agaggcgtcg ggctgctgaa caccgggagc tactatgagc acgtgaagat ttctgcacct
     2161 aatgaatttg atgtcatgtt taaactggaa gtccccagaa ttcaactaga agaatattcc
     2221 aacactcgtg catattactt tgtgaaattt aaaagaaatc cgaaagaaaa tcctctgagt
     2281 cagtttttag aaggtgaaat attatcagct tctaagatgc tgtcaaagtt taggaaaatc
     2341 attaaggaag aaattaacga cattaaagat acagatgtca tcatgaagag gaaaagagga
     2401 gggagccctg ctgtaacact tcttattagt gaaaaaatat ctgtggatat aaccctggct
     2461 ttggaatcaa aaagtagctg gcctgctagc acccaagaag gcctgcgcat tcaaaactgg
     2521 ctttcagcaa aagttaggaa gcaactacga ctaaagccat tttaccttgt acccaagcat
     2581 gcaaaggaag gaaatggttt ccaagaagaa acatggcggc tatccttctc tcacatcgaa
     2641 aaggaaattt tgaacaatca tggaaaatct aaaacgtgct gtgaaaacaa agaagagaaa
     2701 tgttgcagga aagattgttt aaaactaatg aaataccttt tagaacagct gaaagaaagg
     2761 tttaaagaca aaaaacatct ggataaattc tcttcttatc atgtgaaaac tgccttcttt
     2821 cacgtatgta cccagaaccc tcaagacagt cagtgggacc gcaaagacct gggcctctgc
     2881 tttgataact gcgtgacata ctttcttcag tgcctcagga cagaaaaact tgagaattat
     2941 tttattcctg aattcaatct attctctagc aacttaattg acaaaagaag taaagaattt
     3001 ctgacaaagc aaattgaata tgaaagaaac aatgagtttc cagtttttga tgaattttac
     3061 ccatacgatg ttccagatta cgcttgagcg gccgcgccgg cgggcccgaa ttctaccggg
     3121 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     3181 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     3241 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     3301 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     3361 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     3421 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     3481 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     3541 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     3601 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt
     3661 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     3721 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     3781 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3841 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3901 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     3961 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4021 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4081 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4141 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     4201 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg
     4261 cccgaccgaa aggagcgcac gaccccatgc atcggcgtct cgagatatca gtggtccagg
     4321 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     4381 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4441 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     4501 tagagaagtt cagatcaagg tcaggaacag atggaacagg gtcgacccta gagaaccatc
     4561 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     4621 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     4681 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     4741 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     4801 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
     4861 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg
     4921 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4981 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5041 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tagatccggc
     5101 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5161 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5281 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5341 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5401 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttactggc ttaactatgc
     5461 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5521 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6781 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     6841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7081 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     7141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7441 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7501 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt
     7561 agcttggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
     7621 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     7681 ggggtcatta gttcatagcc catatatgga gttccgcgtt a
//
