LOCUS       sequence-366029-        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 cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        332..535
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             536..711
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    774..973
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1174..1590
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1520..1542
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1558..1574
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1630..1979
                     /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    1638..1656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1674..1692
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1710..1728
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1746..1764
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1782..1800
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1818..1836
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1854..1872
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1918..1942
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3084..3110
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3144..3643
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3179..3202)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3576..3595
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3664..4263
                     /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(3664..3683)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4160..4180
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4444..4869
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4972..4994)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5130..5150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5135..5464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5315..5450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5486..5505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5586..5603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5757..6345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5837..5856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6516..7307,7308..7376))
                     /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     7139..7158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7377..7481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7549..7567
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7667..7715,1..331)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
       61 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      121 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      181 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      241 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      301 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      361 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      421 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      481 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata
      541 aaagagccca caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt
      601 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct
      661 tgggagggtc tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc
      721 gtccgggatt tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
      781 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg
      841 cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
      901 ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt
      961 ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg
     1021 gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg
     1081 aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt
     1141 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
     1201 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     1261 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1321 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1381 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1441 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     1501 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     1561 tatccagccc tcactccttc tctaggcgcc ggaattgaag atctcgatga ggccctttcg
     1621 tcttcactcg agtttactcc ctatcagtga tagagaacgt atgaagagtt tactccctat
     1681 cagtgataga gaacgtatgc agactttact ccctatcagt gatagagaac gtataaggag
     1741 tttactccct atcagtgata gagaacgtat gaccagttta ctccctatca gtgatagaga
     1801 acgtatctac agtttactcc ctatcagtga tagagaacgt atatccagtt tactccctat
     1861 cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggcgcctata aaagcagagc
     1921 tcgtttagtg aaccgtcaga tcgcctggag caattccaca acacttttgt cttatacttg
     1981 gatccgccac catgcctggg gcctcgaagc tccgggcggt tttggagaag ttgaagctca
     2041 gccgcgatga tatctccacg gcggcgggga tggtgaaagg ggttgtggac cacctgctgc
     2101 tcagactgaa gtgcgactcc gcgttcagag gcgtcgggct gctgaacacc gggagctact
     2161 atgagcacgt gaagatttct gcacctaatg aatttgatgt catgtttaaa ctggaagtcc
     2221 ccagaattca actagaagaa tattccaaca ctcgtgcata ttactttgtg aaatttaaaa
     2281 gaaatccgaa agaaaatcct ctgagtcagt ttttagaagg tgaaatatta tcagcttcta
     2341 agatgctgtc aaagtttagg aaaatcatta aggaagaaat taacgacatt aaagatacag
     2401 atgtcatcat gaagaggaaa agaggaggga gccctgctgt aacacttctt attagtgaaa
     2461 aaatatctgt ggatataacc ctggctttgg aatcaaaaag tagctggcct gctagcaccc
     2521 aagaaggcct gcgcattcaa aactggcttt cagcaaaagt taggaagcaa ctacgactaa
     2581 agccatttta ccttgtaccc aagcatgcaa aggaaggaaa tggtttccaa gaagaaacat
     2641 ggcggctatc cttctctcac atcgaaaagg aaattttgaa caatcatgga aaatctaaaa
     2701 cgtgctgtga aaacaaagaa gagaaatgtt gcaggaaaga ttgtttaaaa ctaatgaaat
     2761 accttttaga acagctgaaa gaaaggttta aagacaaaaa acatctggat aaattctctt
     2821 cttatcatgt gaaaactgcc ttctttcacg tatgtaccca gaaccctcaa gacagtcagt
     2881 gggaccgcaa agacctgggc ctctgctttg ataactgcgt gacatacttt cttcagtgcc
     2941 tcaggacaga aaaacttgag aattatttta ttcctgaatt caatctattc tctagcaact
     3001 taattgacaa aagaagtaaa gaatttctga caaagcaaat tgaatatgaa agaaacaatg
     3061 agtttccagt ttttgatgaa ttttacccat acgatgttcc agattacgct tgagcggccg
     3121 cgccggcggg cccgaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     3181 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     3241 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     3301 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
     3361 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     3421 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     3481 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     3541 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     3601 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt
     3661 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     3721 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     3781 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     3841 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     3901 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     3961 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4021 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4081 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4141 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4201 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     4261 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     4321 gcgtctcgag atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa
     4381 gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc agaaaaaggg
     4441 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
     4501 ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg
     4561 aacagggtcg accctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     4621 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     4681 tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat
     4741 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     4801 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     4861 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
     4921 caccgggagg taagctggct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4981 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5041 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     5101 tagcgatagc ggagtgtaga tccggctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5281 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5341 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5401 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5461 caaaaagctt actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5521 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     5581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6181 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     6841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7501 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7561 acgaggccct ttcgtcttca agaattagct tggccattgc atacgttgta tccatatcat
     7621 aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg
     7681 actagttatt aatagtaatc aattacgggg tcatt
//
