LOCUS       sequence-366049-        7715 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 7715)
  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 7715)
  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 cGAS:156-522aa-HA
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             535..710
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    773..972
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1173..1589
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1519..1541
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1557..1573
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1629..1978
                     /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    1637..1655
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1673..1691
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1709..1727
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1745..1763
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1781..1799
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1817..1835
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1853..1871
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1917..1941
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3095..3121
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3155..3642
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3190..3213)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3575..3594
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3663..4262
                     /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(3663..3682)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4159..4179
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4443..4868
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4971..4993)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5129..5149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5134..5463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5314..5449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5485..5504
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5585..5602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5756..6344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5836..5855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6515..7306,7307..7375))
                     /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     7138..7157
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7376..7480)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7548..7566
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7666..7715,1..330)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa
      541 aagagcccac aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta
      601 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt
      661 gggagggtct cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg
      721 tccgggattt ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
      781 cagcaactta tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc
      841 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
      901 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
      961 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     1021 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     1081 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     1141 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
     1201 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     1261 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     1321 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1381 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1441 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     1501 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     1561 atccagccct cactccttct ctaggcgccg gaattgaaga tctcgatgag gccctttcgt
     1621 cttcactcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     1681 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     1741 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     1801 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     1861 agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct
     1921 cgtttagtga accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttgg
     1981 atccgccacc atgagggatg cggcgcctgg ggcctcgaag ctccgggcgg ttttggagaa
     2041 gttgaagctc agccgcgatg atatctccac ggcggcgggg atggtgaaag gggttgtgga
     2101 ccacctgctg ctcagactga agtgcgactc cgcgttcaga ggcgtcgggc tgctgaacac
     2161 cgggagctac tatgagcacg tgaagatttc tgcacctaat gaatttgatg tcatgtttaa
     2221 actggaagtc cccagaattc aactagaaga atattccaac actcgtgcat attactttgt
     2281 gaaatttaaa agaaatccga aagaaaatcc tctgagtcag tttttagaag gtgaaatatt
     2341 atcagcttct aagatgctgt caaagtttag gaaaatcatt aaggaagaaa ttaacgacat
     2401 taaagataca gatgtcatca tgaagaggaa aagaggaggg agccctgctg taacacttct
     2461 tattagtgaa aaaatatctg tggatataac cctggctttg gaatcaaaaa gtagctggcc
     2521 tgctagcacc caagaaggcc tgcgcattca aaactggctt tcagcaaaag ttaggaagca
     2581 actacgacta aagccatttt accttgtacc caagcatgca aaggaaggaa atggtttcca
     2641 agaagaaaca tggcggctat ccttctctca catcgaaaag gaaattttga acaatcatgg
     2701 aaaatctaaa acgtgctgtg aaaacaaaga agagaaatgt tgcaggaaag attgtttaaa
     2761 actaatgaaa taccttttag aacagctgaa agaaaggttt aaagacaaaa aacatctgga
     2821 taaattctct tcttatcatg tgaaaactgc cttctttcac gtatgtaccc agaaccctca
     2881 agacagtcag tgggaccgca aagacctggg cctctgcttt gataactgcg tgacatactt
     2941 tcttcagtgc ctcaggacag aaaaacttga gaattatttt attcctgaat tcaatctatt
     3001 ctctagcaac ttaattgaca aaagaagtaa agaatttctg acaaagcaaa ttgaatatga
     3061 aagaaacaat gagtttccag tttttgatga attttaccca tacgatgttc cagattacgc
     3121 ttgagcggcc gcgccggcgg gcccgaattc taccgggtag gggaggcgct tttcccaagg
     3181 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     3241 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     3301 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     3361 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     3421 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     3481 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     3541 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     3601 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     3661 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     3721 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     3781 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3841 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3901 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     3961 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4021 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4081 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4141 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4201 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4261 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg
     4321 cgtctcgaga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     4381 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     4441 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4501 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     4561 acagggtcga ccctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     4621 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     4681 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     4741 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     4801 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     4861 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
     4921 accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4981 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5041 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     5101 agcgatagcg gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5281 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5341 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5401 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5461 aaaaagctta ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5521 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     5581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6181 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6361 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6601 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6661 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6721 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6781 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     6841 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6901 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6961 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7021 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7081 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     7141 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7201 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7261 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7321 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7381 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7441 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7501 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7561 cgaggccctt tcgtcttcaa gaattagctt ggccattgca tacgttgtat ccatatcata
     7621 atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga
     7681 ctagttatta atagtaatca attacggggt catta
//
