LOCUS       sequence-366081-        8144 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Doxycycline-dependent expression of mouse cGAS gene in mammalian 
            cells by retroviral transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  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 8144)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-mouse cGAS-HA
FEATURES             Location/Qualifiers
     source          1..8144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        351..554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             555..730
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    793..992
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1193..1609
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1539..1561
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1577..1593
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1649..1998
                     /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    1657..1675
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1693..1711
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1729..1747
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1765..1783
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1801..1819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1837..1855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1873..1891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1937..1961
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2005..2014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3532..3558
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3592..4091
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3627..3650)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4024..4043
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4112..4711
                     /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(4112..4131)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4608..4628
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4892..5317
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5420..5442)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5578..5598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5583..5912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5763..5898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5934..5953
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6034..6051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6205..6793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6285..6304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6964..7755,7756..7824))
                     /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     7587..7606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7825..7929)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7997..8015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8115..8144,1..350)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
       61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      541 atataagcag agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcttccga
      601 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa
      661 tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cacgacgggg
      721 gtctttcatt tgggggctcg tccgggattt ggagacccct gcccagggac caccgaccca
      781 ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat
      841 gtttgatgtt atgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac
      901 ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga
      961 ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc
     1021 gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     1081 atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat
     1141 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1201 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1261 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1321 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1381 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1441 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1501 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1561 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg gaattgaaga
     1621 tctcgatgag gccctttcgt cttcactcga gtttactccc tatcagtgat agagaacgta
     1681 tgaagagttt actccctatc agtgatagag aacgtatgca gactttactc cctatcagtg
     1741 atagagaacg tataaggagt ttactcccta tcagtgatag agaacgtatg accagtttac
     1801 tccctatcag tgatagagaa cgtatctaca gtttactccc tatcagtgat agagaacgta
     1861 tatccagttt actccctatc agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg
     1921 gcgcctataa aagcagagct cgtttagtga accgtcagat cgcctggagc aattccacaa
     1981 cacttttgtc ttatacttgg atccgccacc atggaagatc cgcgtagaag gacgacggcg
     2041 ccacgcgcta agaagccgtc cgcgaagcgc gccccgacgc agcccagcag gaccagggcc
     2101 cacgcggaaa gctgcggccc gcaaaggggg gctcgatcgc ggcgggcgga gcgtgacggg
     2161 gacaccacgg agaagccacg tgccccaggg ccccgagtgc atccagcaag ggccactgag
     2221 ctcaccaaag atgcacagcc ctcggccatg gacgcggcag gagccaccgc gcggcctgcc
     2281 gtccgggtgc cccagcagca ggccatcctg gatccggagc tgcccgccgt acgggagccc
     2341 cagccgcccg cggatcccga ggcgcggaaa gtcgtaaggg gacctagcca cagaaggggc
     2401 gcgcgctcca ccgggcagcc cagagcgccg cgagggtccg ggaaggaacc ggacaagcta
     2461 aagaaggtgc tggacaaatt gagattgaaa cgcaaagata tctcggaggc ggccgagacg
     2521 gtgaataaag ttgtggaacg cctgctgcgc agaatgcaga aacgggagtc ggagttcaaa
     2581 ggtgtggagc agctgaacac tggcagctac tatgaacatg tgaagatttc tgctcctaat
     2641 gaatttgatg ttatgtttaa actggaagtc cccaggattg agctacaaga atattatgaa
     2701 acaggtgctt tctatcttgt gaaattcaaa agaattccac gaggaaatcc gctgagtcat
     2761 ttcttagaag gggaagtatt atcagctacc aagatgctgt caaagtttag gaaaatcatt
     2821 aaagaagaag ttaaagaaat caaagatata gatgtcagtg tggagaagga aaaaccagga
     2881 agccctgctg taacacttct tatcaggaac cctgaagaaa tctctgtgga tataattctg
     2941 gctttggagt caaaaggcag ctggcctatt agtaccaaag aaggactacc tattcaaggc
     3001 tggctgggca caaaagtgag gaccaatcta agacgagagc cgttttatct cgtacccaag
     3061 aatgcaaagg atggaaatag ttttcaagga gagacctggc gcctctcttt ctctcacact
     3121 gaaaagtaca ttttgaataa tcacgggata gagaaaacat gctgtgaatc ttccggagca
     3181 aaatgctgca gaaaagaatg tttaaaatta atgaaatacc ttttggaaca gttgaaaaaa
     3241 gagtttcaag agctggatgc attctgttcc taccatgtga aaactgccat ctttcacatg
     3301 tggacccagg acccgcagga cagtcagtgg gaccccagga acctcagctc ctgcttcgat
     3361 aagttgttag cattctttct tgagtgcctc aggacagaga aactggatca ttattttatt
     3421 ccaaagttca atctattctc tcaagaacta attgaccgaa aaagtaaaga atttctatcg
     3481 aagaaaattg aatatgaaag aaataatggg tttccaattt ttgacaagct ttacccatac
     3541 gatgttccag attacgcttg agcggccgcg ccggcgggcc cgaattctac cgggtagggg
     3601 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
     3661 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
     3721 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc
     3781 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta
     3841 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
     3901 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg
     3961 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
     4021 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
     4081 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct
     4141 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4201 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     4261 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4321 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4381 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4441 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4501 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4561 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4621 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4681 ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac
     4741 cgaaaggagc gcacgacccc atgcatcggc gtctcgagat atcagtggtc caggctctag
     4801 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     4861 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     4921 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     4981 agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac catcagatgt
     5041 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     5101 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     5161 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     5221 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     5281 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     5341 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     5401 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5461 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5521 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc cggctgtgga
     5581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5821 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5881 gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact atgcggcatc
     5941 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6001 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6061 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6121 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6181 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6241 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6301 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6361 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6421 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6481 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6541 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6601 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6661 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6721 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6781 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6841 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6901 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6961 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7021 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7081 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7141 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7201 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7261 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7321 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7381 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7441 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7501 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7561 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     7621 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7681 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7741 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7801 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7861 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7921 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7981 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattagcttg
     8041 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     8101 attaccgcca tgttgacatt gattattgac tagttattaa tagt
//
