LOCUS       sequence-366575-        7765 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 7765)
  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 7765)
  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 cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        330..533
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             534..709
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    772..971
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1172..1588
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1518..1540
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1556..1572
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1628..1977
                     /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    1636..1654
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1672..1690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1708..1726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1744..1762
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1780..1798
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1816..1834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1852..1870
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1916..1940
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3082..3108
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3177..3200)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     gap             3527..3578
                     /estimated_length=52
     primer_bind     3624..3643
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3712..4311
                     /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(3712..3731)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4208..4228
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4492..4917
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5020..5042)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5178..5198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5183..5512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5363..5498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5534..5553
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5634..5651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5805..6393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5885..5904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6564..7355,7356..7424))
                     /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     7187..7206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7425..7529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7597..7615
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7715..7765,1..329)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
       61 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      121 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      181 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      241 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      301 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      361 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      421 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      481 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcaataaa
      541 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      601 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      661 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      721 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      781 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      841 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      901 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      961 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
     1021 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
     1081 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1141 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1201 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1261 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1321 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1381 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1441 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1501 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1561 tccagccctc actccttctc taggcgccgg aattgaagat ctcgatgagg ccctttcgtc
     1621 ttcactcgag tttactccct atcagtgata gagaacgtat gaagagttta ctccctatca
     1681 gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt ataaggagtt
     1741 tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt gatagagaac
     1801 gtatctacag tttactccct atcagtgata gagaacgtat atccagttta ctccctatca
     1861 gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg cgcctataaa agcagagctc
     1921 gtttagtgaa ccgtcagatc gcctggagca attccacaac acttttgtct tatacttgga
     1981 tccgccacca tgcctggggc caggaagctc cgggcggttt tggagaagtt aaggctcagc
     2041 caccaggaca tctccaaggc ggcgaaggtg gtgaatgggg tagtgggcca cctgctgcgc
     2101 agattgaagt gcgagcccga gttcggaggc gtcgagcagc tgcacaccgg gagctactac
     2161 gagcacgtga agatttctgc acctaatgaa tttgatgtca tgtttaaact gcaagtcccc
     2221 agaatccagc tagaagaata ttctgacact ggtgcatatt actttgtgaa atttaaaaga
     2281 aatccagaag gaaatcctct gagcctgttt ttagaagatg aaatattatc agcttctaag
     2341 atgctgtcaa agtttaggaa aatcattaag gaagaaatta acaacattaa agatgcagat
     2401 gtcatcatga agaggaaaag accagggagc cctgctgtaa cacttcttat taataaagaa
     2461 atatctgtgg atataaccct ggctttggaa ttaaaaacta gctggcctgc cagcacccaa
     2521 gaaggcctgg ccattaaaaa ctggctttca gcaaaagtta ggcagcaact aagactaaag
     2581 ccgttttacc ttgtacccaa gcatgcaaag gaaggaaatg gtttccaaga ggaaacatgg
     2641 cggctatcct tctctcacat cgaaaaggaa attttgagca atcatggaaa atctaaaacg
     2701 tgctgtgaaa acaaagaaat gaaatgttgc aggaaagatt gtttaaaact aatgaaatac
     2761 cttttagaac agctgaaaga aaagtttaaa gacaaaaaac atctggataa attctcttct
     2821 tatcatgtga aaactgcctt ctttcaccta tgtaccgaga accctcaaga cagtcagtgg
     2881 gaccccaaag acctgggcct ctgctttgat aactgcgtga catactttct tgagtgcctc
     2941 aggacagaac aacttaagaa ttattttatt cctggattca atctattctc tagcaactta
     3001 attgacaaaa caagtaaaga atttctgtca aagcaaattg aatatgaaag aaacaatgag
     3061 tttccagttt ttggtgaatt ttacccatac gatgttccag attacgcttg agcggccgcg
     3121 ccggcgggcc cgaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     3181 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     3241 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     3301 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     3361 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     3421 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     3481 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggnnnn nnnnnnnnnn
     3541 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnks skcrggggcg gggcgggcgc
     3601 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     3661 gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac catgaccgag
     3721 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3781 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3841 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3901 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3961 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4021 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4081 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4141 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4201 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4261 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     4321 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcggc gtctcgagat
     4381 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta
     4441 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     4501 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     4561 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagggtcgac
     4621 cctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     4681 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     4741 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
     4801 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     4861 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
     4921 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta
     4981 agctggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5041 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5101 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     5161 agtgtagatc cggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     5221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     5281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     5521 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     5581 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     5641 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5701 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5761 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5821 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5881 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5941 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6001 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6061 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6121 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6181 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6241 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6301 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6361 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6421 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6481 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6541 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6601 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6661 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6721 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6781 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6841 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     6901 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6961 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7021 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7081 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7141 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     7201 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7261 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7321 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7381 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7441 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7501 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7561 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7621 cgtcttcaag aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt
     7681 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
     7741 tagtaatcaa ttacggggtc attag
//
