LOCUS       sequence-366042-        7688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-dependent expression of human cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7688)
  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 7688)
  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-human cGASdeltaN
FEATURES             Location/Qualifiers
     source          1..7688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        294..497
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             498..673
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    736..935
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1136..1552
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1482..1504
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1520..1536
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1592..1941
                     /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    1600..1618
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     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"
     primer_bind     1880..1904
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        3079..3578
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3114..3137)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3511..3530
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3599..4198
                     /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(3599..3618)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4095..4115
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4379..4804
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4907..4929)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5065..5085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5070..5399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5250..5385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5421..5440
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5521..5538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5692..6280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5772..5791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6451..7242,7243..7311))
                     /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     7074..7093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7312..7416)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7484..7502
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7602..7688,1..293)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
       61 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      181 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      301 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      361 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      421 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      481 tctatataag cagagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcttc
      541 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
      601 gaatcgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccacgacg
      661 ggggtctttc atttgggggc tcgtccggga tttggagacc cctgcccagg gaccaccgac
      721 ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc
      781 tatgtttgat gttatgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
      841 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
      901 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
      961 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     1021 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     1081 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     1141 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     1201 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     1261 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     1321 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     1381 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     1441 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     1501 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattga
     1561 agatctcgat gaggcccttt cgtcttcact cgagtttact ccctatcagt gatagagaac
     1621 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca
     1681 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt
     1741 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac
     1801 gtatatccag tttactccct atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg
     1861 tgggcgccta taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg agcaattcca
     1921 caacactttt gtcttatact tggatccgcc accatgcctg gggcctcgaa gctccgggcg
     1981 gttttggaga agttgaagct cagccgcgat gatatctcca cggcggcggg gatggtgaaa
     2041 ggggttgtgg accacctgct gctcagactg aagtgcgact ccgcgttcag aggcgtcggg
     2101 ctgctgaaca ccgggagcta ctatgagcac gtgaagattt ctgcacctaa tgaatttgat
     2161 gtcatgttta aactggaagt ccccagaatt caactagaag aatattccaa cactcgtgca
     2221 tattactttg tgaaatttaa aagaaatccg aaagaaaatc ctctgagtca gtttttagaa
     2281 ggtgaaatat tatcagcttc taagatgctg tcaaagttta ggaaaatcat taaggaagaa
     2341 attaacgaca ttaaagatac agatgtcatc atgaagagga aaagaggagg gagccctgct
     2401 gtaacacttc ttattagtga aaaaatatct gtggatataa ccctggcttt ggaatcaaaa
     2461 agtagctggc ctgctagcac ccaagaaggc ctgcgcattc aaaactggct ttcagcaaaa
     2521 gttaggaagc aactacgact aaagccattt taccttgtac ccaagcatgc aaaggaagga
     2581 aatggtttcc aagaagaaac atggcggcta tccttctctc acatcgaaaa ggaaattttg
     2641 aacaatcatg gaaaatctaa aacgtgctgt gaaaacaaag aagagaaatg ttgcaggaaa
     2701 gattgtttaa aactaatgaa atacctttta gaacagctga aagaaaggtt taaagacaaa
     2761 aaacatctgg ataaattctc ttcttatcat gtgaaaactg ccttctttca cgtatgtacc
     2821 cagaaccctc aagacagtca gtgggaccgc aaagacctgg gcctctgctt tgataactgc
     2881 gtgacatact ttcttcagtg cctcaggaca gaaaaacttg agaattattt tattcctgaa
     2941 ttcaatctat tctctagcaa cttaattgac aaaagaagta aagaatttct gacaaagcaa
     3001 attgaatatg aaagaaacaa tgagtttcca gtttttgatg aattttgagc ggccgcgccg
     3061 gcgggcccga attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     3121 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     3181 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     3241 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     3301 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     3361 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     3421 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     3481 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     3541 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     3601 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3661 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     3721 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     3781 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     3841 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3901 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     3961 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     4021 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     4081 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     4141 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     4201 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggcgtc
     4261 tcgagatatc agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta
     4321 tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4381 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     4441 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     4501 ggtcgaccct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     4561 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     4621 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     4681 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     4741 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     4801 catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg
     4861 ggaggtaagc tggctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4921 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4981 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     5041 atagcggagt gtagatccgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5101 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5221 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5281 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5341 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5401 agcttactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5461 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     5521 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5581 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5881 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6121 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6421 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6481 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6541 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6601 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6661 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6721 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     6781 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6841 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6901 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6961 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7021 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     7081 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7141 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7201 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7261 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7381 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7441 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7501 gccctttcgt cttcaagaat tagcttggcc attgcatacg ttgtatccat atcataatat
     7561 gtacatttat attggctcat gtccaacatt accgccatgt tgacattgat tattgactag
     7621 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     7681 tacataac
//
