LOCUS       sequence-366074-        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of rhesus macaque cGAS gene in 
            mammalian cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  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 7703)
  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-rhesus macaque cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             510..685
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    748..947
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1148..1564
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1494..1516
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1532..1548
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1604..1953
                     /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    1612..1630
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1648..1666
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1684..1702
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1720..1738
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1756..1774
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1792..1810
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1828..1846
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1892..1916
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3058..3084
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3118..3605
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3153..3176)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3538..3557
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3626..4225
                     /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(3626..3645)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4122..4142
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4406..4831
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4934..4956)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5092..5112)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5097..5426
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5277..5412
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5448..5467
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5548..5565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5719..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5799..5818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..7269,7270..7338))
                     /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     7101..7120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7339..7443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7511..7529
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7629..7703,1..305)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctc aataaaagag cccacaaccc ctcactcggc
      541 gcgccagtct tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg
      601 ctgtttgcat ccgaatcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
      661 ctacccacga cgggggtctt tcatttgggg gctcgtccgg gatttggaga cccctgccca
      721 gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga
      781 ttgtctagtg tctatgtttg atgttatgcg cctgcgtctg tactagttag ctaactagct
      841 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
      901 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
      961 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
     1021 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
     1081 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1141 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     1201 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     1261 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     1321 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     1381 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     1441 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     1501 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     1561 cgccggaatt gaagatctcg atgaggccct ttcgtcttca ctcgagttta ctccctatca
     1621 gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt
     1681 tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac
     1741 gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca
     1801 gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt atgtcgaggt
     1861 aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct
     1921 ggagcaattc cacaacactt ttgtcttata cttggatccg ccaccatgcc tggggccagg
     1981 aagctccggg cggttttgga gaagttaagg ctcagccacc aggacatctc caaggcggcg
     2041 aaggtggtga atggggttgt gggccacctg ctgcgcagat tgaagtgcga gcccgagttc
     2101 ggaggcgtcg agcagctgca caccgggagc tactacgagc acgtgaagat ttctgcacct
     2161 aatgaattcg atgtcatgtt taaactgcaa gtccccagaa ttcagctaga agaatattct
     2221 gacactggtg catattactt tgtgaaattt aaaagaaatc cagaaggaaa tcctctgagc
     2281 ctgtttttag aagatgaaat attatcagct tctaagatgc tgtcaaagtt taggaaaatc
     2341 attaaggaag aaattaacaa cattaaagat gcagatgtca tcatgaagag gaaaagacca
     2401 gggagccctg ctgtaacact tcttattaat aaagaaatat ctgtggatat aaccctggct
     2461 ttggaattaa aaactagctg gcctgccagc acccaagaag gcctggccat taaaaactgg
     2521 ctttcagcaa aagttaggca gcaactaaga ctaaagccgt tttaccttgt acccaagcat
     2581 gcaaaggaag gaaatggttt ccaagaggaa acatggcggc tatccttctc tcacatcgaa
     2641 aaggaaattt tgagcaatca tggaaaatct aaaacgtgct gtgaaaacaa agaaatgaaa
     2701 tgttgcagga aagattgttt aaaactaatg aaataccttt tagaacagct gaaagaaaag
     2761 tttaaagaca aaaaacatct ggataaattc tcttcttatc atgtgaaaac tgccttcttt
     2821 cacctatgta ccgagaaccc tcaagacagt cagtgggacc ccaaagacct gggcctctgc
     2881 tttgataact gcgtgacata ctttcttgag tgcctcagga cagaacaact taagaattat
     2941 tttattcctg gattcaatct attctctagc aacttaattg acaaaacaag taaagaattt
     3001 ctgtcaaagc aaattgaata tgaaagaaac aaggagtttc cagtttttgg tgaattttac
     3061 ccatacgatg ttccagatta cgcttgagcg gccgcgccgg cgggcccgaa ttctaccggg
     3121 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     3181 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     3241 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     3301 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     3361 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     3421 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     3481 ggtccggggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     3541 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
     3601 tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     3661 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3721 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3781 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3841 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3901 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3961 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4021 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4081 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4141 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4201 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     4261 gagcgcacga ccccatgcat cggcgtctcg agatatcagt ggtccaggct ctagttttga
     4321 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     4381 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4441 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4501 gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag atgtttccag
     4561 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     4621 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     4681 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     4741 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     4801 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     4861 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg
     4921 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4981 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5041 gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt
     5101 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5161 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5221 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5281 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5341 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5401 tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg catcagagca
     5461 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5521 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6121 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7501 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag cttggccatt
     7561 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     7621 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7681 tcatagccca tatatggagt tcc
//
