LOCUS       sequence-366039-        7715 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 7715)
  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 7715)
  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 cGASdeltaN C396/397A-HA
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        316..519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             520..695
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    758..957
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1158..1574
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1504..1526
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1542..1558
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1614..1963
                     /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    1622..1640
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1658..1676
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1694..1712
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1730..1748
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1766..1784
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1802..1820
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1838..1856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1902..1926
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3068..3094
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3128..3627
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3163..3186)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3560..3579
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3648..4247
                     /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(3648..3667)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4144..4164
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4428..4853
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4956..4978)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5114..5134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5119..5448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5299..5434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5470..5489
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5570..5587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5741..6329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5821..5840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6500..7291,7292..7360))
                     /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     7123..7142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7361..7465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7533..7551
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7651..7715,1..315)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
       61 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      121 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      181 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      241 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      301 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      361 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      421 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      481 ggtaggcgtg tacggtggga ggtctatata agcagagctc aataaaagag cccacaaccc
      541 ctcactcggc gcgccagtct tccgatagac tgcgtcgccc gggtacccgt attcccaata
      601 aagcctcttg ctgtttgcat ccgaatcgtg gtctcgctgt tccttgggag ggtctcctct
      661 gagtgattga ctacccacga cgggggtctt tcatttgggg gctcgtccgg gatttggaga
      721 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt
      781 gtctgtccga ttgtctagtg tctatgtttg atgttatgcg cctgcgtctg tactagttag
      841 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc
      901 gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa
      961 gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt
     1081 cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt
     1141 atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact
     1201 ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg
     1261 ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct
     1321 ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac
     1381 acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct
     1441 gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc
     1501 tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc
     1561 cttctctagg cgccggaatt gaagatctcg atgaggccct ttcgtcttca ctcgagttta
     1621 ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt
     1681 atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt
     1741 gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta
     1801 ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt
     1861 atgtcgaggt aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt
     1921 cagatcgcct ggagcaattc cacaacactt ttgtcttata cttggatccg ccaccatgcc
     1981 tggggcctcg aagctccggg cggttttgga gaagttgaag ctcagccgcg atgatatctc
     2041 cacggcggcg gggatggtga aaggggttgt ggaccacctg ctgctcagac tgaagtgcga
     2101 ctccgcgttc agaggcgtcg ggctgctgaa caccgggagc tactatgagc acgtgaagat
     2161 ttctgcacct aatgaatttg atgtcatgtt taaactggaa gtccccagaa ttcaactaga
     2221 agaatattcc aacactcgtg catattactt tgtgaaattt aaaagaaatc cgaaagaaaa
     2281 tcctctgagt cagtttttag aaggtgaaat attatcagct tctaagatgc tgtcaaagtt
     2341 taggaaaatc attaaggaag aaattaacga cattaaagat acagatgtca tcatgaagag
     2401 gaaaagagga gggagccctg ctgtaacact tcttattagt gaaaaaatat ctgtggatat
     2461 aaccctggct ttggaatcaa aaagtagctg gcctgctagc acccaagaag gcctgcgcat
     2521 tcaaaactgg ctttcagcaa aagttaggaa gcaactacga ctaaagccat tttaccttgt
     2581 acccaagcat gcaaaggaag gaaatggttt ccaagaagaa acatggcggc tatccttctc
     2641 tcacatcgaa aaggaaattt tgaacaatca tggaaaatct aaaacggccg ctgaaaacaa
     2701 agaagagaaa tgttgcagga aagattgttt aaaactaatg aaataccttt tagaacagct
     2761 gaaagaaagg tttaaagaca aaaaacatct ggataaattc tcttcttatc atgtgaaaac
     2821 tgccttcttt cacgtatgta cccagaaccc tcaagacagt cagtgggacc gcaaagacct
     2881 gggcctctgc tttgataact gcgtgacata ctttcttcag tgcctcagga cagaaaaact
     2941 tgagaattat tttattcctg aattcaatct attctctagc aacttaattg acaaaagaag
     3001 taaagaattt ctgacaaagc aaattgaata tgaaagaaac aatgagtttc cagtttttga
     3061 tgaattttac ccatacgatg ttccagatta cgcttgagcg gccgcgccgg cgggcccgaa
     3121 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     3181 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     3241 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     3301 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     3361 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     3421 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     3481 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     3541 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     3601 tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca
     3661 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3721 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3781 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3841 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3901 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3961 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4021 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4081 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4141 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4201 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     4261 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcggcgtct cgagatatca
     4321 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     4381 ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     4441 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     4501 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagg gtcgacccta
     4561 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4621 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     4681 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     4741 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     4801 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
     4861 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
     4921 ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4981 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5041 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     5101 tagatccggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5161 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5221 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5281 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5341 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5401 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttactggc
     5461 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     5521 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     5581 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5641 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5701 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5761 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5821 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5881 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5941 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6001 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6061 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6121 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6181 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6241 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6301 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6361 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6421 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6481 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6541 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6601 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6661 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6721 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6781 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     6841 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6901 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6961 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7021 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7081 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     7141 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7201 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7261 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7321 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7381 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7441 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     7501 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     7561 ttcaagaatt agcttggcca ttgcatacgt tgtatccata tcataatatg tacatttata
     7621 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
     7681 aatcaattac ggggtcatta gttcatagcc catat
//
