LOCUS       sequence-366441-        7787 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 7787)
  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 7787)
  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-ER
FEATURES             Location/Qualifiers
     source          1..7787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        324..527
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             528..703
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    766..965
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1166..1582
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1512..1534
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1550..1566
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1622..1971
                     /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    1630..1648
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1666..1684
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1702..1720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1738..1756
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1774..1792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1810..1828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1846..1864
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1910..1934
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3208..3707
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3243..3266)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3640..3659
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3728..4327
                     /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(3728..3747)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4224..4244
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4508..4933
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5036..5058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5194..5214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5199..5528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5379..5514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5550..5569
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5650..5667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5821..6409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5901..5920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6580..7371,7372..7440))
                     /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     7203..7222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7441..7545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7613..7631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7731..7787,1..323)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
       61 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      361 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      421 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      481 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcaa taaaagagcc
      541 cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg gtacccgtat
      601 tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg
      661 tctcctctga gtgattgact acccacgacg ggggtctttc atttgggggc tcgtccggga
      721 tttggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac
      781 ttatctgtgt ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc tgcgtctgta
      841 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac
      901 acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac
      961 ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga
     1021 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag
     1081 ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt
     1141 gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt
     1201 aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag
     1261 acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga
     1321 cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc
     1381 gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc
     1441 ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc
     1501 cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc
     1561 cctcactcct tctctaggcg ccggaattga agatctcgat gaggcccttt cgtcttcact
     1621 cgagtttact ccctatcagt gatagagaac gtatgaagag tttactccct atcagtgata
     1681 gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg agtttactcc
     1741 ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga gaacgtatct
     1801 acagtttact ccctatcagt gatagagaac gtatatccag tttactccct atcagtgata
     1861 gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag
     1921 tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatact tggatccgcc
     1981 accatgcctg gggcctcgaa gctccgggcg gttttggaga agttgaagct cagccgcgat
     2041 gatatctcca cggcggcggg gatggtgaaa ggggttgtgg accacctgct gctcagactg
     2101 aagtgcgact ccgcgttcag aggcgtcggg ctgctgaaca ccgggagcta ctatgagcac
     2161 gtgaagattt ctgcacctaa tgaatttgat gtcatgttta aactggaagt ccccagaatt
     2221 caactagaag aatattccaa cactcgtgca tattactttg tgaaatttaa aagaaatccg
     2281 aaagaaaatc ctctgagtca gtttttagaa ggtgaaatat tatcagcttc taagatgctg
     2341 tcaaagttta ggaaaatcat taaggaagaa attaacgaca ttaaagatac agatgtcatc
     2401 atgaagagga aaagaggagg gagccctgct gtaacacttc ttattagtga aaaaatatct
     2461 gtggatataa ccctggcttt ggaatcaaaa agtagctggc ctgctagcac ccaagaaggc
     2521 ctgcgcattc aaaactggct ttcagcaaaa gttaggaagc aactacgact aaagccattt
     2581 taccttgtac ccaagcatgc aaaggaagga aatggtttcc aagaagaaac atggcggcta
     2641 tccttctctc acatcgaaaa ggaaattttg aacaatcatg gaaaatctaa aacgtgctgt
     2701 gaaaacaaag aagagaaatg ttgcaggaaa gattgtttaa aactaatgaa atacctttta
     2761 gaacagctga aagaaaggtt taaagacaaa aaacatctgg ataaattctc ttcttatcat
     2821 gtgaaaactg ccttctttca cgtatgtacc cagaaccctc aagacagtca gtgggaccgc
     2881 aaagacctgg gcctctgctt tgataactgc gtgacatact ttcttcagtg cctcaggaca
     2941 gaaaaacttg agaattattt tattcctgaa ttcaatctat tctctagcaa cttaattgac
     3001 aaaagaagta aggaatttct gacaaagcaa attgaatatg aaagaaacaa tgagtttcca
     3061 gtttttgatg aatttctcaa gcgcaaatac tggtggaaaa acctcaagat gatgatcatc
     3121 ttgggagtga tttgcgccat catcctcatc atcatcatag tttacttcag cacttaagcg
     3181 gccgcgccgg cgggcccgaa ttctaccggg taggggaggc gcttttccca aggcagtctg
     3241 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     3301 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     3361 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     3421 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     3481 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     3541 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     3601 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     3661 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa
     3721 gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     3781 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3841 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3901 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3961 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4021 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4081 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4141 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4201 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4261 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4321 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc
     4381 atcggcgtct cgagatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc
     4441 tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa
     4501 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     4561 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     4621 atggaacagg gtcgacccta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     4681 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     4741 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     4801 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     4861 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     4921 ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac
     4981 ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     5041 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5101 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     5161 cacgtagcga tagcggagtg tagatccggc tgtggaatgt gtgtcagtta gggtgtggaa
     5221 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5281 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5341 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5401 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5461 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5521 tttgcaaaaa gcttactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5581 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     5641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6241 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     6901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     7201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7561 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     7621 tatcacgagg ccctttcgtc ttcaagaatt agcttggcca ttgcatacgt tgtatccata
     7681 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt
     7741 attgactagt tattaatagt aatcaattac ggggtcatta gttcata
//
