LOCUS       sequence-366041-        8150 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 8150)
  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 8150)
  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
FEATURES             Location/Qualifiers
     source          1..8150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        326..529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             530..705
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    768..967
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1168..1584
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1514..1536
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1552..1568
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1624..1973
                     /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    1632..1650
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1668..1686
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1704..1722
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1740..1758
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1776..1794
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1812..1830
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1848..1866
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1912..1936
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3585..4072
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3620..3643)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4005..4024
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4093..4692
                     /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(4093..4112)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4589..4609
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4873..5298
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5401..5423)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5559..5579)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5564..5893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5744..5879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5915..5934
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6015..6032)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6186..6774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6266..6285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6945..7736,7737..7805))
                     /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     7568..7587
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7806..7910)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7978..7996
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8096..8150,1..325)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
       61 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      121 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      181 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      301 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc aataaaagag
      541 cccacaaccc ctcactcggc gcgccagtct tccgatagac tgcgtcgccc gggtacccgt
      601 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gtctcgctgt tccttgggag
      661 ggtctcctct gagtgattga ctacccacga cgggggtctt tcatttgggg gctcgtccgg
      721 gatttggaga cccctgccca gggaccaccg acccaccacc gggaggtaag ctggccagca
      781 acttatctgt gtctgtccga ttgtctagtg tctatgtttg atgttatgcg cctgcgtctg
      841 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      901 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      961 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
     1021 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
     1081 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
     1141 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1201 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1261 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1321 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1381 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1441 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1501 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1561 gccctcactc cttctctagg cgccggaatt gaagatctcg atgaggccct ttcgtcttca
     1621 ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga
     1681 tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact
     1741 ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat
     1801 ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga
     1861 tagagaacgt atgtcgaggt aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt
     1921 agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata cttggatccg
     1981 ccaccatgca gccttggcac ggaaaggcca tgcagagagc ttccgaggcc ggagccactg
     2041 cccccaaggc ttccgcacgg aatgccaggg gcgccccgat ggatcccacc gagtctccgg
     2101 ctgcccccga ggccgccctg cctaaggcgg gaaagttcgg ccccgccagg aagtcgggat
     2161 cccggcagaa aaagagcgcc ccggacaccc aggagaggcc gcccgtccgc gcaactgggg
     2221 cccgcgccaa aaaggcccct cagcgcgccc aggacacgca gccgtctgac gccaccagcg
     2281 cccctggggc agaggggctg gagcctcctg cggctcggga gccggctctt tccagggctg
     2341 gttcttgccg ccagaggggc gcgcgctgct ccacgaagcc aagacctccg cccgggccct
     2401 gggacgtgcc cagccccggc ctgccggtct cggcccccat tctcgtacgg agggatgcgg
     2461 cgcctggggc ctcgaagctc cgggcggttt tggagaagtt gaagctcagc cgcgatgata
     2521 tctccacggc ggcggggatg gtgaaagggg ttgtggacca cctgctgctc agactgaagt
     2581 gcgactccgc gttcagaggc gtcgggctgc tgaacaccgg gagctactat gagcacgtga
     2641 agatttctgc acctaatgaa tttgatgtca tgtttaaact ggaagtcccc agaattcaac
     2701 tagaagaata ttccaacact cgtgcatatt actttgtgaa atttaaaaga aatccgaaag
     2761 aaaatcctct gagtcagttt ttagaaggtg aaatattatc agcttctaag atgctgtcaa
     2821 agtttaggaa aatcattaag gaagaaatta acgacattaa agatacagat gtcatcatga
     2881 agaggaaaag aggagggagc cctgctgtaa cacttcttat tagtgaaaaa atatctgtgg
     2941 atataaccct ggctttggaa tcaaaaagta gctggcctgc tagcacccaa gaaggcctgc
     3001 gcattcaaaa ctggctttca gcaaaagtta ggaagcaact acgactaaag ccattttacc
     3061 ttgtacccaa gcatgcaaag gaaggaaatg gtttccaaga agaaacatgg cggctatcct
     3121 tctctcacat cgaaaaggaa attttgaaca atcatggaaa atctaaaacg tgctgtgaaa
     3181 acaaagaaga gaaatgttgc aggaaagatt gtttaaaact aatgaaatac cttttagaac
     3241 agctgaaaga aaggtttaaa gacaaaaaac atctggataa attctcttct tatcatgtga
     3301 aaactgcctt ctttcacgta tgtacccaga accctcaaga cagtcagtgg gaccgcaaag
     3361 acctgggcct ctgctttgat aactgcgtga catactttct tcagtgcctc aggacagaaa
     3421 aacttgagaa ttattttatt cctgaattca atctattctc tagcaactta attgacaaaa
     3481 gaagtaaaga atttctgaca aagcaaattg aatatgaaag aaacaatgag tttccagttt
     3541 ttgatgaatt ttgagcggcc gcgccggcgg gcccgaattc taccgggtag gggaggcgct
     3601 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     3661 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     3721 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     3781 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     3841 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     3901 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     3961 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     4021 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     4081 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4141 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4201 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4261 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4321 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4381 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4441 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4501 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4561 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4621 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4681 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     4741 catgcatcgg cgtctcgaga tatcagtggt ccaggctcta gttttgactc aacaatatca
     4801 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     4861 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     4921 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     4981 aacagatgga acagggtcga ccctagagaa ccatcagatg tttccagggt gccccaagga
     5041 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5101 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     5161 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     5221 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     5281 cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca
     5341 ccgacccacc accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa
     5401 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5461 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     5521 ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg
     5581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5641 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5821 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5881 aggcttttgc aaaaagctta ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5941 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6001 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6061 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6601 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6901 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6961 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7021 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7081 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7141 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7201 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7261 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7321 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7381 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7441 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7501 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7561 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7621 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7681 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7741 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7801 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7861 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7921 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7981 aggcgtatca cgaggccctt tcgtcttcaa gaattagctt ggccattgca tacgttgtat
     8041 ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat
     8101 tgattattga ctagttatta atagtaatca attacggggt cattagttca
//
