LOCUS       sequence-366437-        7631 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 7631)
  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 7631)
  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-HA-human cGASdeltaN
FEATURES             Location/Qualifiers
     source          1..7631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        336..539
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             540..715
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    778..977
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1178..1594
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1524..1546
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1562..1578
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1634..1983
                     /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    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"
     protein_bind    1858..1876
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1922..1946
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1999..2025
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3148..3635
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3183..3206)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3568..3587
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3656..4255
                     /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(3656..3675)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4152..4172
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4436..4861
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4964..4986)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5122..5142)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5166..5376
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5188..5384
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5235..5370
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5406..5425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5506..5523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5677..6265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5757..5776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6436..7227,7228..7296))
                     /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     7059..7078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7297..7401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7469..7487
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7587..7631,1..335)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
       61 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      121 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      181 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      241 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      301 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      361 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      421 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      481 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      541 aataaaagag cccacaaccc ctcactcggc gcgccagtct tccgatagac tgcgtcgccc
      601 gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gtctcgctgt
      661 tccttgggag ggtctcctct gagtgattga ctacccacga cgggggtctt tcatttgggg
      721 gctcgtccgg gatttggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
      781 ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgtttg atgttatgcg
      841 cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga
      901 cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt
      961 ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt
     1021 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
     1081 ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct
     1141 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
     1201 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
     1261 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
     1321 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
     1381 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
     1441 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
     1501 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
     1561 cctttatcca gccctcactc cttctctagg cgccggaatt gaagatctcg atgaggccct
     1621 ttcgtcttca ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc
     1681 ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa
     1741 ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata
     1801 gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc
     1861 ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggcgcc tataaaagca
     1921 gagctcgttt agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata
     1981 cttggatccg ccaccatgta cccatacgat gttccagatt acgctcctgg ggcctcgaag
     2041 ctccgggcgg ttttggagaa gttgaagctc agccgcgatg atatctccac ggcggcgggg
     2101 atggtgaaag gggttgtgga ccacctgctg ctcagactga agtgcgactc cgcgttcaga
     2161 ggcgtcgggc tgctgaacac cgggagctac tatgagcacg tgaagatttc tgcacctaat
     2221 gaatttgatg tcatgtttaa actggaagtc cccagaattc aactagaaga atattccaac
     2281 actcgtgcat attactttgt gaaatttaaa agaaatccga aagaaaatcc tctgagtcag
     2341 tttttagaag gtgaaatatt atcagcttct aagatgctgt caaagtttag gaaaatcatt
     2401 aaggaagaaa ttaacgacat taaagataca gatgtcatca tgaagaggaa aagaggaggg
     2461 agccctgctg taacacttct tattagtgaa aaaatatctg tggatataac cctggctttg
     2521 gaatcaaaaa gtagctggcc tgctagcacc caagaaggcc tgcgcattca aaactggctt
     2581 tcagcaaaag ttaggaagca actacgacta aagccatttt accttgtacc caagcatgca
     2641 aaggaaggaa atggtttcca agaagaaaca tggcggctat ccttctctca catcgaaaag
     2701 gaaattttga acaatcatgg aaaatctaaa acgtgctgtg aaaacaaaga agagaaatgt
     2761 tgcaggaaag attgtttaaa actaatgaaa taccttttag aacagctgaa agaaaggttt
     2821 aaagacaaaa aacatctgga taaattctct tcttatcatg tgaaaactgc cttctttcac
     2881 gtatgtaccc agaaccctca agacagtcag tgggaccgca aagacctggg cctctgcttt
     2941 gataactgcg tgacatactt tcttcagtgc ctcaggacag aaaaacttga gaattatttt
     3001 attcctgaat tcaatctatt ctctagcaac ttaattgaca aaagaagtaa agaatttctg
     3061 acaaagcaaa ttgaatatga aagaaacaat gagtttccag tttttgatga attttgagcg
     3121 gccgcgccgg cgggcccgaa ttctaccggg taggggaggc gcttttccca aggcagtctg
     3181 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     3241 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     3301 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     3361 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     3421 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     3481 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcggggcgg
     3541 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg
     3601 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac
     3661 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     3721 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     3781 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     3841 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     3901 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     3961 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4021 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4081 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4141 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4201 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc
     4261 gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cggcgtctcg
     4321 agatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag
     4381 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4441 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4501 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagggt
     4561 cgaccctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4621 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4681 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     4741 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4801 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     4861 ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga
     4921 ggtaagctgg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4981 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5041 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     5101 gcggagtgta gatccggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     5161 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5221 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5281 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5341 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt actggcttaa
     5401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     5461 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     5521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6721 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     6781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7021 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     7081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7441 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     7501 agaattagct tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
     7561 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     7621 aattacgggg t
//
