LOCUS       sequence-366068-        8165 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 8165)
  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 8165)
  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-gibbon cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8165
                     /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"
     CDS             3526..3552
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3586..4085
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3621..3644)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4018..4037
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4106..4705
                     /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(4106..4125)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4602..4622
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4886..5311
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5414..5436)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5572..5592)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5577..5906
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5757..5892
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5928..5947
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6028..6045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6199..6787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6279..6298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6958..7749,7750..7818))
                     /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     7581..7600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7819..7923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7991..8009
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8109..8165,1..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 accatgcgga gagattccga ggccggagcc actgcctccg aggcttccgc gcggagtgcc
     2041 aggggcgccc cgacggaccc caacgagtct ccggctgccc ccgaggccgc cctgcctaag
     2101 gcgggaaagt tcggccccgc caggaagtcg ggtagtcggc agaaaaagag cgccccggac
     2161 acccaggaga ggccgcccgt ccgcgcaact ggggctcgcg ccaaaaaggc ccctcagcgc
     2221 gcccaggacg cgcagccctc tgacgccacc agcgcccctg gggcagaggg gctggagcct
     2281 cctgcggctc gggagccggc tctttccagg gctggttctc gccaccagag aggcgcgcgc
     2341 tgctctccga agccaagacc cccgcccggg cccggggacg tgcccagccc cggcctgccg
     2401 gtctcggacc ccattctcgt acagggagat gcggcgcctg gggccaggaa gctccgggcg
     2461 gttttggaga agttgaagct cagccgccag gaaatctccg aggcggcgga ggtggtgaac
     2521 ggggttgtgg accacctgct gcgcagactg cagaagtgcg actccgagtt cagaggcgtc
     2581 gggctgctgc gcaccgggag ctactatgag cacgtgaaga tttctgcacc taatgaattt
     2641 gacgtcatgt ttaaactgga agtccccaga atccaactag aagaatattc caacactggt
     2701 gcatattact ttgtgaaatt taaaagaaat ccgaaagaaa atcctttgag tcagttttta
     2761 gaaggtgaaa tattatcagc ttctaagatg ctgtcaaaat ttaggaaaat cattaaggaa
     2821 gaaattaaca acattaaaga tacagatgtc atcatgaaga ggaaaagagg agggagccct
     2881 gctgtaacac ttcttattag tgaaaaaata tctgtggata taaccctggc tttggaatca
     2941 aacagtaata gctggcctgc tagcacccaa gaaggcctgc ccattaaaga ctggctttca
     3001 ggaaaagtta ggaggcaact aagactaaag ccattttacc ttgtacccaa gcatgcaaag
     3061 gaaggaaatg gtttccaaga agaaacatgg cggctatcct tctctcacat cgaaaaggaa
     3121 attttgaaca atcatggaaa aactaaaacg tgctgtgaaa acaaaaatgt gaaatgttgc
     3181 aggaaagatt gtttaaaact aatgaaatac cttttagaac agctgaaaga aaggtttaaa
     3241 gacaaaaaac atctggataa attctcttct tatcatgtga aaactgcctt cttccatgta
     3301 tgtaccgagt accctcaaga cagtcagtgg gaccccaaaa acctgggctt ctgctttgat
     3361 aaatgcgtga tatactttct taagtgcctc aggacagaac aacttgggaa ttattttatt
     3421 cctggattca atctattctc tagcaactta attgacaaaa gaagtaaaga atttctgtca
     3481 aagcaaattg aatatgaaag aaacaatgag tatccagttt ttggttaccc atacgatgtt
     3541 ccagattacg cttgagcggc cgcgccggcg ggcccgaatt ctaccgggta ggggaggcgc
     3601 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
     3661 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
     3721 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
     3781 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
     3841 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
     3901 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
     3961 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     4021 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
     4081 tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     4141 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4201 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4261 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4321 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4381 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4441 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4501 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4561 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4621 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4681 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     4741 gagcgcacga ccccatgcat cggcgtctcg agatatcagt ggtccaggct ctagttttga
     4801 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     4861 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4921 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4981 gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag atgtttccag
     5041 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     5101 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     5161 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     5221 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     5281 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     5341 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg
     5401 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5461 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5521 gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt
     5581 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5641 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5701 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5761 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5821 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5881 tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg catcagagca
     5941 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6001 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6061 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6601 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7261 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7561 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7921 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7981 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag cttggccatt
     8041 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     8101 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     8161 tcata
//
