LOCUS       sequence-366079-        7706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of orangutan cGAS gene in mammalian
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7706)
  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 7706)
  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-orangutan cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7706
                     /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             3087..3113
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3147..3634
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3182..3205)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3567..3586
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3655..4254
                     /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(3655..3674)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4151..4171
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4435..4860
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4963..4985)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5121..5141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5126..5455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5306..5441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5477..5496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5577..5594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5748..6336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5828..5847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6507..7298,7299..7367))
                     /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     7130..7149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7368..7472)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7540..7558
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7658..7706,1..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 gccgcctgga aatctccaag gcggcggagg tggtgaacag ggttgtggac cacctgctgc
     2101 gcagactgca gaagtacaac tccgagttca gaggcgtcgg gctgctgcgc accgggagct
     2161 actatgagca cgtgaagatt tctgcaccta atgaatttga tgtcatgttt aaactggaag
     2221 tccccagaat tcaactagaa gaatattcca acactggtgc atattacttt gtgaaattta
     2281 aaagaaatcc gaaagaaaat cctctgagtc agtttttaga aggtgaaata ttatcagctt
     2341 ctaagatgct gtcaaagttt aggaaaatca ttaaggaaga aattaacaac attaaagata
     2401 cagatgtcat catgaagagg aaaagaggag ggagccctgc tgtaacactt cttattagtg
     2461 aagaaatatc tgtggatata accctggctt tggaatcaaa aagtagctgg cctgctagca
     2521 cccaagaagg cctgcccatt aaaaactggc tttcagcaaa agttaggaag caactaagac
     2581 taaagccatt ttaccttgta cccaagcatg caaaggaagg aaatggtttc caagaagaaa
     2641 catggcggct atccttctct cacatcgaaa aggaaatttt gaacaatcat ggaaaatcta
     2701 aaacgtgctg tgaagacaaa gaagtgaaat gttgcaggaa agattgttta aaactaatga
     2761 aatacctttt agaacagctg aaagaaaggt ttaaagacaa aaaacatctg gataaattct
     2821 cttcttatca tgtgaaaact gccttctttc acgtatgtac ccagaaccct caagacagtc
     2881 agtgggaccc caaagacctg ggcctctgct ttgataactg cgtgacatac tttcttcagt
     2941 gcctcaggac agaacaactt gggaattatt ttattcctgg attcaatcta ttctctagca
     3001 acttaattga caaaagaagt aaagaatttc tgacaaagca aattgaatat gaaagaaaca
     3061 atgagtttcc agtttttgat gaattttacc catacgatgt tccagattac gcttgagcgg
     3121 ccgcgccggc gggcccgaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     3181 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     3241 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     3301 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     3361 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     3421 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     3481 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcggggcggg
     3541 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     3601 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc
     3661 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3721 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3781 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3841 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3901 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3961 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     4021 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     4081 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     4141 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     4201 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     4261 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc ggcgtctcga
     4321 gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga
     4381 gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     4441 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     4501 aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc
     4561 gaccctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4621 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4681 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     4741 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4801 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     4861 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
     4921 gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4981 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5041 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     5101 cggagtgtag atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5221 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5281 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5341 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5401 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5461 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5521 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     5581 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5641 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5701 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5761 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5821 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5881 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5941 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6001 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6061 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6121 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6181 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6241 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6301 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6361 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6421 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6481 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6541 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6601 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6661 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6721 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6781 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     6841 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6901 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6961 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7021 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7081 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     7141 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7201 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7261 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7321 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7381 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7441 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7501 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7561 tttcgtcttc aagaattagc ttggccattg catacgttgt atccatatca taatatgtac
     7621 atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat
     7681 taatagtaat caattacggg gtcatt
//
