LOCUS       sequence-366071-        7790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of marmoset cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7790)
  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 7790)
  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-marmoset cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        346..549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             550..725
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    788..987
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1188..1604
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1534..1556
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1572..1588
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1644..1993
                     /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    1652..1670
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1688..1706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1724..1742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1760..1778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1796..1814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1832..1850
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1868..1886
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1932..1956
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3173..3199
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3233..3732
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3268..3291)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3665..3684
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3753..4352
                     /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(3753..3772)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4249..4269
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4533..4958
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5061..5083)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5219..5239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5224..5553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5404..5539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5575..5594
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5675..5692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5846..6434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5926..5945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6605..7396,7397..7465))
                     /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     7228..7247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7466..7570)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7638..7656
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7756..7790,1..345)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
       61 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      121 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      181 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      241 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      301 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      361 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      421 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      481 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      541 agcagagctc aataaaagag cccacaaccc ctcactcggc gcgccagtct tccgatagac
      601 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
      661 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccacga cgggggtctt
      721 tcatttgggg gctcgtccgg gatttggaga cccctgccca gggaccaccg acccaccacc
      781 gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgtttg
      841 atgttatgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
      901 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
      961 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     1021 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     1081 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     1141 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     1201 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     1261 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     1321 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     1381 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     1441 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     1501 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     1561 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt gaagatctcg
     1621 atgaggccct ttcgtcttca ctcgagttta ctccctatca gtgatagaga acgtatgaag
     1681 agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga
     1741 gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct
     1801 atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc
     1861 agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggcgcc
     1921 tataaaagca gagctcgttt agtgaaccgt cagatcgcct ggagcaattc cacaacactt
     1981 ttgtcttata cttggatccg ccaccatgcc gcggaccagg aagctccggg cggttttaga
     2041 gaagttaagg ctaagtcgcg acgatgtctc caacgcggcc aaggtggtga acagcgttgt
     2101 ggaccacctg ctgggcagac tgcagcagtg cgagtccgag ttcaaaggca agttcaaagt
     2161 ggagtcgctg cacaccggga gctactatga gcgcgtgaag atttctacac ctaatgaatt
     2221 tgatgtcatg tttaaactgg aactccccag aatccaacta gaagaatatt ccaacactgg
     2281 tgcatattac tttgtgaaat tcaaaagaaa tctgaaagaa aatcctctga gacaattttt
     2341 agaagatgaa atattatcag cttctaagat gctgtctaag tttaggaaaa tcattaagga
     2401 agaaattaaa aacatgcaag atacagatgt catcatggag aggaaaagaa gagggagccc
     2461 tgctgtaaca cttcttatta gaaaagaaat atctgtggat ataatcctgg ctttggaatc
     2521 aaaaagtagc tggcctgcta gcacccagaa aggcctgccc atcaaaaact ggctttcagc
     2581 gaaagttagg acgaagctaa gacgagagcc attttacctt gtacccaagc atgcaaagga
     2641 aggaaatagt ttccaagaag aaacatggcg gctatccttc tctcacattg aaaaggaaat
     2701 tttgagggat catggacaat ctaaaacgtg ttgtgaagag aaaggagtga aatgttgcag
     2761 gaaagattgt ttaaaactga tgaaatacct tttagaacgg ctgaaagaaa agtttaaaag
     2821 tggaaaacat atggataaat tctgttctta tcatgtgaaa actgctttct ttcacgtatg
     2881 taccgagaac cctcaagaca gtcagtggga ccccaaagat ctggagctct gctttgataa
     2941 ctgcctgaca tactttcttc agtgcctcag gacagaaaaa cttgaacatt atttcattcc
     3001 tgcattcaat ctattctcta gcaacttaat tgacaaaaaa agtaaagaat ttctgtcaaa
     3061 gcaaattgaa catgaaagaa acaatgagtt tccagttttt ggtgaatttc gagattctat
     3121 gtttagaaaa aatttgagtg actctcagtc ctggagaata ccaaaaaaat tttacccata
     3181 cgatgttcca gattacgctt gagcggccgc gccggcgggc ccgaattcta ccgggtaggg
     3241 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     3301 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     3361 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     3421 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     3481 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     3541 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     3601 gggggcgggc tcaggggnng gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     3661 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     3721 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     3781 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     3841 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     3901 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     3961 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4021 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     4081 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     4141 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     4201 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     4261 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     4321 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     4381 ccgaaaggag cgcacgaccc catgcatcgg cgtctcgaga tatcagtggt ccaggctcta
     4441 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     4501 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     4561 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     4621 aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa ccatcagatg
     4681 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     4741 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     4801 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     4861 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     4921 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg
     4981 agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg cctcgcgcgt
     5041 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5101 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5161 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg
     5221 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5281 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5341 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5401 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5461 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5521 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat
     5581 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     5641 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6421 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6481 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6541 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6601 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6781 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6901 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7021 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7201 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     7261 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7441 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7501 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7561 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7621 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattagctt
     7681 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa
     7741 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca
//
