LOCUS       sequence-366077-        8177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of crab-eating 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 8177)
  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 8177)
  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-crab-eating macaque cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        329..532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             533..708
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    771..970
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1171..1587
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1517..1539
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1555..1571
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1627..1976
                     /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    1635..1653
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1671..1689
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1707..1725
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1743..1761
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1779..1797
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1815..1833
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1851..1869
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1915..1939
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3555..3581
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3615..4102
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3650..3673)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4035..4054
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4123..4722
                     /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(4123..4142)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4619..4639
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4903..5328
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5431..5453)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5589..5609)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5594..5923
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5774..5909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5945..5964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6045..6062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6216..6804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6296..6315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6975..7766,7767..7835))
                     /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     7598..7617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7836..7940)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8008..8026
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8126..8177,1..328)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
       61 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      121 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      181 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcaataaaa
      541 gagcccacaa cccctcactc ggcgcgccag tcttccgata gactgcgtcg cccgggtacc
      601 cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggtctcgc tgttccttgg
      661 gagggtctcc tctgagtgat tgactaccca cgacgggggt ctttcatttg ggggctcgtc
      721 cgggatttgg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggcca
      781 gcaacttatc tgtgtctgtc cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt
      841 ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
      901 tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc
      961 ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta
     1021 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac
     1081 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg
     1141 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg
     1201 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
     1261 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
     1321 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
     1381 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
     1441 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca
     1501 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat
     1561 ccagccctca ctccttctct aggcgccgga attgaagatc tcgatgaggc cctttcgtct
     1621 tcactcgagt ttactcccta tcagtgatag agaacgtatg aagagtttac tccctatcag
     1681 tgatagagaa cgtatgcaga ctttactccc tatcagtgat agagaacgta taaggagttt
     1741 actccctatc agtgatagag aacgtatgac cagtttactc cctatcagtg atagagaacg
     1801 tatctacagt ttactcccta tcagtgatag agaacgtata tccagtttac tccctatcag
     1861 tgatagagaa cgtatgtcga ggtaggcgtg tacggtgggc gcctataaaa gcagagctcg
     1921 tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt atacttggat
     1981 ccgccaccat gcagcctcgg caaggaaagg ccacgcggag agcttccgag gccggagcca
     2041 ctgtccccga ggcttccgcg cggagtgcca ggggggcccg gacggaccgc aacgagtctc
     2101 cggctgcccc cgaagccgcc ctgcccgagg cgggaaagtt cggccccgcc aggaagtcgg
     2161 gttcagggca gaagaagagc cccccggaca cccaggagcg gtcgccagtc cgcgcaactg
     2221 gggcccgcgc taaaaaggcc cctcagcgcg cccaggacgc gcggccctct gactccacca
     2281 gcgccgctgg ggcggaggtg ctggagcctc ctgcggctcg ggaaccggct ctttccaggg
     2341 ctggttctcg ccgccagagg ggcgcgcgct gctcttcgaa gccaagaccc ccgcccgggc
     2401 cccgggatgt gcccaacccc ggcctgccgg tctcggtccc cattctggta catagggatg
     2461 cggcgcctgg ggccaggaag ctccgggcgg ttttggagaa gttaaggctc agccaccagg
     2521 acatctccaa ggcggcgaag gtggtgaatg gggtagtggg ccacctgctg cgcagattga
     2581 agtgcgagcc cgagttcgga ggcgtcgagc agctgcacac cgggagctac tacgagcacg
     2641 tgaagatttc tgcacctaat gaatttgatg tcatgtttaa actgcaagtc cccagaatcc
     2701 agctagaaga atattctgac actggtgcat attactttgt gaaatttaaa agaaatccag
     2761 aaggaaatcc tctgagcctg tttttagaag atgaaatatt atcagcttct aagatgctgt
     2821 caaagtttag gaaaatcatt aaggaagaaa ttaacaacat taaagatgca gatgtcatca
     2881 tgaagaggaa aagaccaggg agccctgctg taacacttct tattaataaa gaaatatctg
     2941 tggatataac cctggctttg gaattaaaaa ctagctggcc tgccagcacc caagaaggcc
     3001 tggccattaa aaactggctt tcagcaaaag ttaggcagca actaagacta aagccgtttt
     3061 accttgtacc caagcatgca aaggaaggaa atggtttcca agaggaaaca tggcggctat
     3121 ccttctctca catcgaaaag gaaattttga gcaatcatgg aaaatctaaa acgtgctgtg
     3181 aaaacaaaga aatgaaatgt tgcaggaaag attgtttaaa actaatgaaa taccttttag
     3241 aacagctgaa agaaaagttt aaagacaaaa aacatctgga taaattctct tcttatcatg
     3301 tgaaaactgc cttctttcac ctatgtaccg agaaccctca agacagtcag tgggacccca
     3361 aagacctggg cctctgcttt gataactgcg tgacatactt tcttgagtgc ctcaggacag
     3421 aacaacttaa gaattatttt attcctggat tcaatctatt ctctagcaac ttaattgaca
     3481 aaacaagtaa agaatttctg tcaaagcaaa ttgaatatga aagaaacaat gagtttccag
     3541 tttttggtga attttaccca tacgatgttc cagattacgc ttgagcggcc gcgccggcgg
     3601 gcccgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     3661 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     3721 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     3781 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     3841 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     3901 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     3961 gaggctggga aggggtgggt ccgggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     4021 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     4081 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     4141 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4201 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4261 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4321 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4381 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4441 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4501 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4561 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4621 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4681 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     4741 cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg cgtctcgaga tatcagtggt
     4801 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     4861 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     4921 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     4981 gagaatagag aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa
     5041 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     5101 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     5161 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     5221 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     5281 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc
     5341 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg
     5401 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5461 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5521 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat
     5581 ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5641 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5701 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5761 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5821 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5881 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac
     5941 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6001 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     6061 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7261 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     7321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7561 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     7621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7681 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7981 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     8041 gaattagctt ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc
     8101 tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca
     8161 attacggggt cattagt
//
