LOCUS       sequence-366075-        8294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of white-handed gibbon cGAS gene in
            mammalian cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8294)
  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 8294)
  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-WHG cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8294
                     /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"
     CDS             3656..3682
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3716..4215
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3751..3774)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4148..4167
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4236..4835
                     /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(4236..4255)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4732..4752
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5016..5441
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5544..5566)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5702..5722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5707..6036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5887..6022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6058..6077
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6158..6175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6329..6917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6409..6428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7088..7879,7880..7948))
                     /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     7711..7730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7949..8053)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8121..8139
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8239..8294,1..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 caccatgcgg agagattccg aggccggagc cactgcctcc gaggcttccg cgcggagtgc
     2041 caggggcgcc ccgacggatc ccaacgagtc tccggctgct ccagaggccg ccctgcctaa
     2101 ggcgggaaag ttcggccccg ccaggaagtc gggatcccgg cagaaaaaga gcgccccgga
     2161 cacccaggag aggccgcccg tccgcgcaac tggggctcgc gccaaaaagg cccctcagcg
     2221 cgcccaggac gcgcagccct ctgacgccac cagcgcccct ggggcagagg ggctggagcc
     2281 tcctgcggct cgggagccgg ctctttccag ggctggttct cgcgccaaaa aggcccctca
     2341 gcgctcccag gacgcgcagc cctctgacgc caccagcgcc cctggggcag aggggctgga
     2401 gcctcctgcg gctcgggagc tggctctttc cagggctggt tcccgccacc agagaggcgc
     2461 gcgctgctct ccgaagccaa gacccccgcc cgggcccggg gacgtgccca gccccggcat
     2521 gccggtctcg gcccccattc tcgtacaggg ggatgcggcg cctggggcca ggaagctccc
     2581 ggcggttttg gagaagttga agctcagccg ccaggaaatc tgcgaggcgg cggaggtggt
     2641 gaacggggtt gtggaccacc tgctgcgcag actgcagaag tgcgactccg agttcagagg
     2701 cgtcgggctg ctgcgcaccg ggagctacta tgagcacgtg aagatttctg cacctaatga
     2761 atttgacgtc atgtttaaac tggaagtccc cagaattcaa ctagaagaat attccaacac
     2821 tggtgcatat tactttgtga aatttaaaag aaatccgaaa gaaaatcctc tgtgtcagtt
     2881 tttagaaggt gaaatattat cagcttctaa gatgctgtca aagtttagga aaatcattaa
     2941 ggaagaaatt aacaacatta aagatacaga tgtcatcctg aagaggaaaa gaggagggag
     3001 ccctgctgta acacttctta ttagtgaaaa aatatctgtg gatataaccc tggctttgga
     3061 atcaaagagt agtagctggc ctgctagcac ccaagaaggc ctgcccatta aagactggct
     3121 ttcaggaaaa gttaggacgc aactaagact aaagccattt taccttgtac ccaagcatgc
     3181 aaaggaagga aatggtttcc aagaagaaac atggcggcta tccttctctc acattgaaaa
     3241 ggaaattttg aacaatcatg gagaaactaa aacgtgctgt gaaaacaaaa aagtgaaatg
     3301 ttgcaggaaa gattgtttaa aactaatgaa atacctttta gaacagctga aagaaaggtt
     3361 taaagacaaa aaacatctgg ataaattctc ttcttatcat gtgaaaactg ccttctttca
     3421 tgtatgtacc gagaacccaa aagacagtca gtgggacccc aaagacctgg gcctctgctt
     3481 tgataaatgc gtgatatact ttcttcagtg cctcagtaca gaacaacttg ggaattattt
     3541 tattcctgga ttcaatctat tctctagcaa cttaattgac aaaagaagta aagaatttct
     3601 gtcaaagcaa attgaatatg aaagaaacaa tgagtatcca gtttttggta aatgttaccc
     3661 atacgatgtt ccagattacg cttgagcggc cgcgccggcg ggcccgaatt ctaccgggta
     3721 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     3781 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     3841 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     3901 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     3961 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     4021 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     4081 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     4141 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     4201 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     4261 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4321 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4381 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4441 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4501 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4561 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4621 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4681 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4741 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4801 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     4861 cgaccgaaag gagcgcacga ccccatgcat cggcgtctcg agatatcagt ggtccaggct
     4921 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     4981 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     5041 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     5101 gagaagttca gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag
     5161 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     5221 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     5281 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     5341 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     5401 cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc
     5461 gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg
     5521 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5581 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5641 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg
     5701 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5761 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5821 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5881 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5941 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6001 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg
     6061 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6121 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6181 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6241 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6301 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6361 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6421 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6481 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6541 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6601 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6661 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6721 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6781 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6841 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6901 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6961 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7021 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7081 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7141 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7201 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7261 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7321 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7381 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7441 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7501 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7561 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7621 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7681 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     7741 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7801 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7861 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7921 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7981 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8041 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     8101 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag
     8161 cttggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     8221 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     8281 ggtcattagt tcat
//
