LOCUS       sequence-366160-        7790 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 7790)
  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 7790)
  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-OMM
FEATURES             Location/Qualifiers
     source          1..7790
                     /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"
     promoter        3215..3702
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3250..3273)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3635..3654
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3723..4322
                     /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(3723..3742)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4219..4239
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4503..4928
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5031..5053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5189..5209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5194..5523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5374..5509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5545..5564
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5645..5662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5816..6404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5896..5915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6575..7366,7367..7435))
                     /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     7198..7217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7436..7540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7608..7626
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7726..7790,1..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 ccacatcgaa aaggaaattt tgaacaatca tggaaaatct aaaacgtgct gtgaaaacaa
     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 tgaatttgag agggaggtgc catgccacag gccctcacct ggggctctgt ggctccaggt
     3121 ggctgtgaca ggggtgctgg tagtcacact cctggtggtg ctgtaccggc ggcgtctgca
     3181 ctaggcggcc gcgccggcgg gcccgaattc taccgggtag gggaggcgct tttcccaagg
     3241 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     3301 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     3361 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     3421 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     3481 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     3541 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     3601 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     3661 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     3721 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     3781 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     3841 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3901 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3961 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4021 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4081 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4141 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4201 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4261 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4321 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg
     4381 cgtctcgaga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     4441 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     4501 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4561 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     4621 acagggtcga ccctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     4681 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     4741 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     4801 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     4861 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     4921 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
     4981 accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     5161 agcgatagcg gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5221 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5281 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5341 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5401 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5461 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5521 aaaaagctta ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5581 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     5641 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5701 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5761 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5821 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5881 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5941 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6001 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6061 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6121 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6181 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6241 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6301 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6361 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6421 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6481 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6541 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6601 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6661 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6721 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6781 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6841 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     6901 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6961 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7021 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7081 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7141 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     7201 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7261 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7321 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7381 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7441 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7501 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7561 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7621 cgaggccctt tcgtcttcaa gaattagctt ggccattgca tacgttgtat ccatatcata
     7681 atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga
     7741 ctagttatta atagtaatca attacggggt cattagttca tagcccatat
//
