LOCUS       sequence-366064-        7685 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 7685)
  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 7685)
  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 cGAS:166-522aa-HA
FEATURES             Location/Qualifiers
     source          1..7685
                     /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             3064..3090
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3124..3611
                     /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     3544..3563
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3632..4231
                     /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(3632..3651)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4128..4148
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4412..4837
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4940..4962)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5098..5118)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5103..5432
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5283..5418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5454..5473
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5554..5571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5725..6313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5805..5824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6484..7275,7276..7344))
                     /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     7107..7126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7345..7449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7517..7535
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7635..7685,1..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 tgcgggcggt tttggagaag ttgaagctca gccgcgatga tatctccacg
     2041 gcggcgggga tggtgaaagg ggttgtggac cacctgctgc tcagactgaa gtgcgactcc
     2101 gcgttcagag gcgtcgggct gctgaacacc gggagctact atgagcacgt gaagatttct
     2161 gcacctaatg aatttgatgt aatgtttaaa ctggaagtcc ccagaattca actagaagaa
     2221 tattccaaca ctcgtgcata ttactttgtg aaatttaaaa gaaatccgaa agaaaatcct
     2281 ctgagtcagt ttttagaagg tgaaatatta tcagcttcta agatgctgtc aaagtttagg
     2341 aaaatcatta aggaagaaat taacgacatt aaagatacag atgtcatcat gaagaggaaa
     2401 agaggaggga gccctgctgt aacacttctt attagtgaaa aaatatctgt ggatataacc
     2461 ctggctttgg aatcaaaaag tagctggcct gctagcaccc aagaaggcct gcgcattcaa
     2521 aactggcttt cagcaaaagt taggaagcaa ctacgactaa agccatttta ccttgtaccc
     2581 aagcatgcaa aggaaggaaa tggtttccaa gaagaaacat ggcggctatc cttctctcac
     2641 atcgaaaagg aaattttgaa caatcatgga aaatctaaaa cgtgctgtga aaacaaagaa
     2701 gagaaatgtt gcaggaaaga ttgtttaaaa ctaatgaaat accttttaga acagctgaaa
     2761 gaaaggttta aagacaaaaa acatctggat aaattctctt cttatcatgt gaaaactgcc
     2821 ttctttcacg tatgtaccca gaaccctcaa gacagtcagt gggaccgcaa agacctgggc
     2881 ctctgctttg ataactgcgt gacatacttt cttcagtgcc tcaggacaga aaaacttgag
     2941 aattatttta ttcctgaatt caatctattc tctagcaact taattgacaa aagaagtaaa
     3001 gaatttctga caaagcaaat tgaatatgaa agaaacaatg agtttccagt ttttgatgaa
     3061 ttttacccat 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 gggcgggcgc ccgaaggtcc tccggaggcc
     3541 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
     3601 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct
     3661 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     3721 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     3781 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     3841 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     3901 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     3961 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4021 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4081 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4141 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4201 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     4261 cgaaaggagc gcacgacccc atgcatcggc gtctcgagat atcagtggtc caggctctag
     4321 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     4381 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     4441 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     4501 agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac catcagatgt
     4561 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     4621 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     4681 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     4741 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     4801 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     4861 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     4921 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4981 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5041 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc cggctgtgga
     5101 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5161 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5221 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5281 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5341 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5401 gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact atgcggcatc
     5461 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5521 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6781 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7081 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7501 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattagcttg
     7561 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     7621 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     7681 attag
//
