LOCUS       sequence-366080-        7697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of chimpanzee cGAS gene in 
            mammalian cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7697)
  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 7697)
  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-chimpanzee cGASdeltaN-HA
FEATURES             Location/Qualifiers
     source          1..7697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        334..537
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             538..713
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    776..975
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1176..1592
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1522..1544
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1560..1576
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1632..1981
                     /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    1640..1658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1676..1694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1712..1730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1748..1766
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1784..1802
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1820..1838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1856..1874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1920..1944
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3080..3106
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3140..3627
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3175..3198)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3560..3579
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3648..4247
                     /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(3648..3667)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4144..4164
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4428..4853
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4956..4978)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5114..5134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5119..5448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5299..5434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5470..5489
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5570..5587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5741..6329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5821..5840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6500..7291,7292..7360))
                     /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     7123..7142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7361..7465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7533..7551
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7651..7697,1..333)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
       61 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      121 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      181 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      241 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      301 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      361 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      421 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      481 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcaa
      541 taaaagagcc cacaacccct cactcggcgc gccagtcttc cgatagactg cgtcgcccgg
      601 gtacccgtat tcccaataaa gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc
      661 cttgggaggg tctcctctga gtgattgact acccacgacg ggggtctttc atttgggggc
      721 tcgtccggga tttggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
      781 ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc
      841 tgcgtctgta ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg
      901 agttctgaac acccggccgc aaccctggga gacgtcccag ggactttggg ggccgttttt
      961 gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac cccgtcagga tatgtggttc
     1021 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
     1081 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
     1141 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
     1201 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1261 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1321 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1381 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1441 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1501 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1561 tttatccagc cctcactcct tctctaggcg ccggaattga agatctcgat gaggcccttt
     1621 cgtcttcact cgagtttact ccctatcagt gatagagaac gtatgaagag tttactccct
     1681 atcagtgata gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg
     1741 agtttactcc ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga
     1801 gaacgtatct acagtttact ccctatcagt gatagagaac gtatatccag tttactccct
     1861 atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg tgggcgccta taaaagcaga
     1921 gctcgtttag tgaaccgtca gatcgcctgg agcaattcca caacactttt gtcttatact
     1981 tggatccgcc accatgcctg gggcctcgaa gctccgggcg gttttggaga agttgaagct
     2041 cagccgcgat gatatctcca cggcggcggg gatggtgaaa ggggttgtgg accacctgct
     2101 gctcagactg aagtgcgact ccgcgttcag aggcgtcggg ctgctgaaca ccgggagcta
     2161 ctatgagcac gtgaagattt ctgcacctaa tgaatttgat gtcatgttta aactggaagt
     2221 ccccagaatc caactagaag aatattccaa cactcgtgca tattactttg tgaaatttaa
     2281 aagaaatccg aaagaaaatc ctctgagtca gtttttagaa ggtgaaatat tatcagcttc
     2341 taagatgctg tcaaagttta ggaaaatcat taaggaagaa attaacaaca ttaaagatgt
     2401 catcatgaag aggaaaagag gagggagccc tgctgtaaca cttcttatta gtgaaaaaat
     2461 atctgtggat ataaccctgg ctttggaatc aaaaagtagc tggcctgcta gcacccaaga
     2521 aggcctgccc attaaaaact ggctttcagc aaaagttagg aagcaactaa gactaaagcc
     2581 attttacctt gtacccaagc atgcaaagga aggaaatggt ttccaagaag aaacatggcg
     2641 gctatccttc tctcacatcg aaaaggaaat tttgaacaat catggaaaat ctaaaacgtg
     2701 ctgtgaaatc aaagaagcga aatgttgcag gaaagattgt ttaaaactaa tgaaatacct
     2761 tttagaacag ctgaaagaaa agtttaaaga caaaaaacat ctggataaat tctcttctta
     2821 tcatgtgaaa actgccttct ttcacgtatg taccgagaac cctcaagaca gtcagtggga
     2881 ccgcaaagac ctgggcctct gctttgataa ctgcgtgaca tactttcttc agtgcctcag
     2941 gatagaaaaa cttgagaatt attttattcc tgaatttaat ctattctcta gcgacttaat
     3001 tgacaaaaga agtaaagaat ttctgacaaa gcaaattgaa tatgaaagaa acaatgagtt
     3061 tccagttttt gatgaatttt acccatacga tgttccagat tacgcttgag cggccgcgcc
     3121 ggcgggcccg aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     3181 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     3241 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     3301 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     3361 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     3421 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     3481 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcggggc gggcgcccga
     3541 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     3601 tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca
     3661 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3721 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3781 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3841 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3901 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3961 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4021 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     4081 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     4141 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     4201 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     4261 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcggcgtct cgagatatca
     4321 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     4381 ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     4441 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     4501 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagg gtcgacccta
     4561 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4621 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     4681 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     4741 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     4801 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
     4861 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
     4921 ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     4981 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5041 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     5101 tagatccggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5161 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5221 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5281 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5341 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5401 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttactggc
     5461 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     5521 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     5581 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5641 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5701 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5761 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5821 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5881 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5941 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6001 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6061 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6121 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6181 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6241 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6301 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6361 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6421 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6481 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6541 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6601 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6661 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6721 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6781 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     6841 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6901 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6961 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7021 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7081 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     7141 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7201 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7261 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7321 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7381 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7441 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     7501 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     7561 ttcaagaatt agcttggcca ttgcatacgt tgtatccata tcataatatg tacatttata
     7621 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
     7681 aatcaattac ggggtca
//
