LOCUS       sequence-393692-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of gibbon cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  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 7715)
  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-gibbon cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        312..515
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             516..691
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    754..953
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1154..1570
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1500..1522
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1538..1554
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1610..1959
                     /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    1618..1636
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1654..1672
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1690..1708
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1726..1744
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1762..1780
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1798..1816
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1834..1852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1898..1922
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3064..3090
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3124..3623
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3159..3182)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3556..3575
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3644..4243
                     /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(3644..3663)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4140..4160
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4424..4849
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4952..4974)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5110..5130)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5115..5444
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5295..5430
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5466..5485
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5566..5583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5737..6325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5817..5836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6496..7287,7288..7356))
                     /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     7119..7138
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7357..7461)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7529..7547
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7647..7715,1..311)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      301 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      361 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      421 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      481 ggcgtgtacg gtgggaggtc tatataagca gagctcaata aaagagccca caacccctca
      541 ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc ccaataaagc
      601 ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc tcctctgagt
      661 gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt tggagacccc
      721 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt atctgtgtct
      781 gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact agttagctaa
      841 ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac ccggccgcaa
      901 ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct gaggaaggga
      961 gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg agaacctaaa
     1021 acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg
     1081 tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1141 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1201 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1261 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1321 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1381 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1441 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1501 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1561 tctaggcgcc ggaattgaag atctcgatga ggccctttcg tcttcactcg agtttactcc
     1621 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
     1681 agactttact ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata
     1741 gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc
     1801 ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga gaacgtatgt
     1861 cgaggtaggc gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga
     1921 tcgcctggag caattccaca acacttttgt cttatacttg gatccgccac catgcctggg
     1981 gccaggaagc tccgggcggt tttggagaag ttgaagctca gccgccagga aatctccgag
     2041 gcggcggagg tggtgaacgg ggttgtggac cacctgctgc gcagactgca gaagtgcgac
     2101 tccgagttca gaggcgtcgg gctgctgcgc accgggagct actatgagca cgtgaagatt
     2161 tctgcaccta atgaatttga cgtcatgttt aaactggaag tccccagaat ccaactagaa
     2221 gaatattcca acactggtgc atattacttt gtgaaattta aaagaaatcc gaaagaaaat
     2281 cctttgagtc agtttttaga aggtgaaata ttatcagctt ctaagatgct gtcaaaattt
     2341 aggaaaatca ttaaggaaga aattaacaac attaaagata cagatgtcat catgaagagg
     2401 aaaagaggag ggagccctgc tgtaacactt cttattagtg aaaaaatatc tgtggatata
     2461 accctggctt tggaatcaaa cagtaatagc tggcctgcta gcacccaaga aggcctgccc
     2521 attaaagact ggctttcagg aaaagttagg aggcaactaa gactaaagcc attttacctt
     2581 gtacccaagc atgcaaagga aggaaatggt ttccaagaag aaacatggcg gctatccttc
     2641 tctcacatcg aaaaggaaat tttgaacaat catggaaaaa ctaaaacgtg ctgtgaaaac
     2701 aaaaatgtga aatgttgcag gaaagattgt ttaaaactaa tgaaatacct tttagaacag
     2761 ctgaaagaaa ggtttaaaga caaaaaacat ctggataaat tctcttctta tcatgtgaaa
     2821 actgccttct tccatgtatg taccgagtac cctcaagaca gtcagtggga ccccaaaaac
     2881 ctgggcttct gctttgataa atgcgtgata tactttctta agtgcctcag gacagaacaa
     2941 cttgggaatt attttattcc tggattcaat ctattctcta gcaacttaat tgacaaaaga
     3001 agtaaagaat ttctgtcaaa gcaaattgaa tatgaaagaa acaatgagta tccagttttt
     3061 ggttacccat acgatgttcc agattacgct tgagcggccg cgccggcggg cccgaattct
     3121 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     3181 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     3241 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     3301 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     3361 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     3421 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     3481 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     3541 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     3601 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc
     3661 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     3721 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     3781 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     3841 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     3901 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     3961 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4021 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4081 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4141 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4201 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     4261 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gcgtctcgag atatcagtgg
     4321 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     4381 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4441 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     4501 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagggtcg accctagaga
     4561 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     4621 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     4681 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     4741 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     4801 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     4861 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggct
     4921 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4981 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5041 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtaga
     5101 tccggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5161 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5221 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5281 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5341 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5401 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt actggcttaa
     5461 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     5521 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     5581 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5641 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5701 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5761 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5821 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5881 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5941 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6001 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6061 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6121 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6181 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6361 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6421 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6481 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6541 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6601 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6661 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6721 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6781 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     6841 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6901 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6961 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7021 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7081 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     7141 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7201 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7261 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7321 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7381 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     7501 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     7561 agaattagct tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg
     7621 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     7681 aattacgggg tcattagttc atagcccata tatgg
//
