LOCUS       sequence-366440-        7982 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 7982)
  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 7982)
  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-Pex
FEATURES             Location/Qualifiers
     source          1..7982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        325..528
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             529..704
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    767..966
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1167..1583
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1513..1535
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1551..1567
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1623..1972
                     /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    1631..1649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1667..1685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1703..1721
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1739..1757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1775..1793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1811..1829
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1847..1865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1911..1935
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3404..3903
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3439..3462)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3836..3855
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3924..4523
                     /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(3924..3943)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4420..4440
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4704..5129
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5232..5254)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5390..5410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5395..5724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5575..5710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5746..5765
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5846..5863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6017..6605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6097..6116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6776..7567,7568..7636))
                     /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     7399..7418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7637..7741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7809..7827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7927..7982,1..324)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
       61 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      121 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      181 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      241 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      301 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      361 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      421 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      481 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc
      541 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
      601 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
      661 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
      721 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      781 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
      841 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      901 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      961 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     1021 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     1081 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     1141 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1201 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1261 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1321 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1381 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1441 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1501 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1561 ccctcactcc ttctctaggc gccggaattg aagatctcga tgaggccctt tcgtcttcac
     1621 tcgagtttac tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat
     1681 agagaacgta tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc
     1741 cctatcagtg atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc
     1801 tacagtttac tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat
     1861 agagaacgta tgtcgaggta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta
     1921 gtgaaccgtc agatcgcctg gagcaattcc acaacacttt tgtcttatac ttggatccgc
     1981 caccatgcct ggggcctcga agctccgggc ggttttggag aagttgaagc tcagccgcga
     2041 tgatatctcc acggcggcgg ggatggtgaa aggggttgtg gaccacctgc tgctcagact
     2101 gaagtgcgac tccgcgttca gaggcgtcgg gctgctgaac accgggagct actatgagca
     2161 cgtgaagatt tctgcaccta atgaatttga tgtcatgttt aaactggaag tccccagaat
     2221 tcaactagaa gaatattcca acactcgtgc atattacttt gtgaaattta aaagaaatcc
     2281 gaaagaaaat cctctgagtc agtttttaga aggtgaaata ttatcagctt ctaagatgct
     2341 gtcaaagttt aggaaaatca ttaaggaaga aattaacgac attaaagata cagatgtcat
     2401 catgaagagg aaaagaggag ggagccctgc tgtaacactt cttattagtg aaaaaatatc
     2461 tgtggatata accctggctt tggaatcaaa aagtagctgg cctgctagca cccaagaagg
     2521 cctgcgcatt caaaactggc tttcagcaaa agttaggaag caactacgac taaagccatt
     2581 ttaccttgta cccaagcatg caaaggaagg aaatggtttc caagaagaaa catggcggct
     2641 atccttctct cacatcgaaa aggaaatttt gaacaatcat ggaaaatcta aaacgtgctg
     2701 tgaaaacaaa gaagagaaat gttgcaggaa agattgttta aaactaatga aatacctttt
     2761 agaacagctg aaagaaaggt ttaaagacaa aaaacatctg gataaattct cttcttatca
     2821 tgtgaaaact gccttctttc acgtatgtac ccagaaccct caagacagtc agtgggaccg
     2881 caaagacctg ggcctctgct ttgataactg cgtgacatac tttcttcagt gcctcaggac
     2941 agaaaaactt gagaattatt ttattcctga attcaatcta ttctctagca acttaattga
     3001 caaaagaagt aaagaatttc tgacaaagca aattgaatat gaaagaaaca atgagtttcc
     3061 agtttttgat gaatttattg tgcatgcatt tgcctctgtc agtatgatga tggatgctac
     3121 cttttcagct gtctataaca gtttcagggc tgtattggat gtagcaaatc acttttcccg
     3181 attgaaaata cactttacaa aagtgttttc agcttttgca ttggttagga ctatacggta
     3241 tctttacaga cggctacagc ggatgttagg tttaagaaga ggctctgaga atgaagacct
     3301 ctgggcagag agtgaaggaa ctgtggcatg ccttggtgct gaggaccgag cagctacctc
     3361 agcaaaatct tgagcggccg cgccggcggg cccgaattct accgggtagg ggaggcgctt
     3421 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     3481 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     3541 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     3601 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     3661 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     3721 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     3781 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     3841 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     3901 tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     3961 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     4021 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     4081 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     4141 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     4201 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     4261 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     4321 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     4381 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     4441 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     4501 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
     4561 gcgcacgacc ccatgcatcg gcgtctcgag atatcagtgg tccaggctct agttttgact
     4621 caacaatatc accagctgaa gcctatagag tacgagccat agataaaata aaagatttta
     4681 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     4741 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     4801 tcaaggtcag gaacagatgg aacagggtcg accctagaga accatcagat gtttccaggg
     4861 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4921 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4981 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     5041 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     5101 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
     5161 cccagggacc accgacccac caccgggagg taagctggct gcctcgcgcg tttcggtgat
     5221 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5281 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5341 gcagccatga cccagtcacg tagcgatagc ggagtgtaga tccggctgtg gaatgtgtgt
     5401 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5461 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5521 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5581 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5641 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5701 tttggaggcc taggcttttg caaaaagctt actggcttaa ctatgcggca tcagagcaga
     5761 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     5821 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6421 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6661 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7081 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7381 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7741 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     7801 cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattagct tggccattgc
     7861 atacgttgta tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc
     7921 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     7981 at
//
