LOCUS       sequence-366570-        7805 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 7805)
  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 7805)
  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 cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        321..524
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             525..700
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    763..962
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1163..1579
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1509..1531
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1547..1563
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1619..1968
                     /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    1627..1645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1663..1681
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1699..1717
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1735..1753
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1771..1789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1807..1825
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1843..1861
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1907..1931
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3079..3105
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3174..3197)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     gap             3529..3547
                     /estimated_length=19
     primer_bind     3655..3674
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3743..4342
                     /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(3743..3762)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4239..4259
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4523..4948
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5051..5073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5209..5229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5214..5543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5394..5529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5565..5584
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5665..5682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5836..6424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5916..5935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6595..7386,7387..7455))
                     /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     7218..7237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7456..7560)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7628..7646
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7746..7805,1..320)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
       61 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      121 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      181 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      241 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      301 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      361 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      421 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      481 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac
      541 aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc
      601 caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct
      661 cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt
      721 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
      781 tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta
      841 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      901 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
      961 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     1021 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     1081 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1141 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1201 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1261 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1321 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1381 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1441 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1501 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1561 cactccttct ctaggcgccg gaattgaaga tctcgatgag gccctttcgt cttcactcga
     1621 gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag
     1681 aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta
     1741 tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca
     1801 gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag
     1861 aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga
     1921 accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttgg atccgccacc
     1981 atgcctgggg ccaggaagct cccggcggtt ttggagaagt tgaagctcag ccgccaggaa
     2041 atctgcgagg cggcggaggt ggtgaacggg gttgtggacc acctgctgcg cagactgcag
     2101 aagtgcgact ccgagttcag aggcgtcggg ctgctgcgca ccgggagcta ctatgagcac
     2161 gtgaagattt ctgcacctaa tgaatttgac gtcatgttta aactggaagt ccccagaatt
     2221 caactagaag aatattccaa cactggtgca tattactttg tgaaatttaa aagaaatccg
     2281 aaagaaaatc ctctgtgtca gtttttagaa ggtgaaatat tatcagcttc taagatgctg
     2341 tcaaagttta ggaaaatcat taaggaagaa attaacaaca ttaaagatac agatgtcatc
     2401 ctgaagagga aaagaggagg gagccctgct gtaacacttc ttattagtga aaaaatatct
     2461 gtggatataa ccctggcttt ggaatcaaag agtagtagct ggcctgctag cacccaagaa
     2521 ggcctgccca ttaaagactg gctttcagga aaagttagga cgcaactaag actaaagcca
     2581 ttttaccttg tacccaagca tgcaaaggaa ggaaatggtt tccaagaaga aacatggcgg
     2641 ctatccttct ctcacattga aaaggaaatt ttgaacaatc atggagaaac taaaacgtgc
     2701 tgtgaaaaca aaaaagtgaa atgttgcagg aaagattgtt taaaactaat gaaatacctt
     2761 ttagaacagc tgaaagaaag gtttaaagac aaaaaacatc tggataaatt ctcttcttat
     2821 catgtgaaaa ctgccttctt tcatgtatgt accgagaacc caaaagacag tcagtgggac
     2881 cccaaagacc tgggcctctg ctttgataaa tgcgtgatat actttcttca gtgcctcagt
     2941 acagaacaac ttgggaatta ttttattcct ggattcaatc tattctctag caacttaatt
     3001 gacaaaagaa gtaaagaatt tctgtcaaag caaattgaat atgaaagaaa caatgagtat
     3061 ccagtttttg gtaaatgtta cccatacgat gttccagatt acgcttgagc ggccgcgccg
     3121 gcgggcccga attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     3181 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     3241 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     3301 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     3361 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     3421 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     3481 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggcnn nnnnnnnnnn
     3541 nnnnnnnscy cktggttttg ggggcnargg ggnggggagg gggbgggccn gggggcgggc
     3601 tcrggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
     3661 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     3721 cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3781 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3841 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3901 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3961 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4021 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4081 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4141 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4201 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4261 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4321 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     4381 cgcacgaccc catgcatcgg cgtctcgaga tatcagtggt ccaggctcta gttttgactc
     4441 aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa aagattttat
     4501 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     4561 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     4621 caaggtcagg aacagatgga acagggtcga ccctagagaa ccatcagatg tttccagggt
     4681 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     4741 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     4801 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     4861 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     4921 gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc
     4981 ccagggacca ccgacccacc accgggaggt aagctggctg cctcgcgcgt ttcggtgatg
     5041 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     5101 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
     5161 cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg aatgtgtgtc
     5221 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5281 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5341 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5401 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5461 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5521 ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat cagagcagat
     5581 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5641 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5701 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5761 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5821 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5881 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5941 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6001 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6061 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6121 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6181 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6241 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6301 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6361 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6421 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6481 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6541 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6601 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6661 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6721 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6781 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6841 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6901 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6961 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7021 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7081 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7141 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7201 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7261 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7321 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7381 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7441 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7501 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7561 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     7621 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattagctt ggccattgca
     7681 tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc
     7741 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     7801 tagcc
//
