LOCUS       sequence-439175-        8106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible expression of Gag in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8106)
  AUTHORS   Landau LM, Chaudhary N, Tien YC, Rogozinska M, Joshi S, Yao C, 
            Crowley J, Hullahalli K, Campbell IW, Waldor MK, Haigis M, Kagan JC
  TITLE     pLxIS-containing domains are biochemically flexible regulators of 
            interferons and metabolism.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2436-2454.e10. doi: 
            10.1016/j.molcel.2024.05.030. Epub 2024 Jun 25.
  PUBMED    38925114
REFERENCE   2  (bases 1 to 8106)
  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"; doi: 
            "10.1016/j.molcel.2024.05.030"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2436-2454.e10"
COMMENT     Sequence Label: pRetroX-TRE3G-Gag
FEATURES             Location/Qualifiers
     source          1..8106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             16..39
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1556..2055
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1591..1614)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1988..2007
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2076..2675
                     /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(2076..2095)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2572..2592
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2856..3281
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(3384..3406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3542..3562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3547..3876
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3727..3862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3898..3917
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3998..4015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4169..4757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4249..4268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4928..5719,5720..5788))
                     /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     5551..5570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5789..5893)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5961..5979
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        6079..6458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6459..6662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6663..6838
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    6901..7100
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7301..7717
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7647..7669
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7685..7701
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        7757..8106
                     /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    7765..7783
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7801..7819
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7837..7855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7873..7891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7909..7927
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7945..7963
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7981..7999
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     8045..8069
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ggatccgcca ccatggacta caaagacgat gacgacaaga tgggagcacg cgcctccgta
       61 cttagcgggg gcgagcttga caggtgggag aaaattagat tgaggccagg agggaagaaa
      121 aaatacaagt tgaagcatat cgtctgggcc agtcgagagt tggagcgctt tgccgttaac
      181 ccaggtcttc ttgagacctc cgaaggatgt cggcagatct tgggtcagct gcagccaagt
      241 cttcaaacag gttctgaaga gctcaggagt ctgtacaata ctgtagccac actgtactgt
      301 gtccaccaga gaattgaaat aaaagatact aaagaagccc ttgacaagat tgaggaggaa
      361 cagaataaga gtaagaagaa ggcccaacaa gcagcagcag acaccggaca ttctaaccag
      421 gtttctcaaa actaccccat agtacagaat attcagggtc aaatggttca ccaggctatc
      481 tccccccgca ccctgaatgc atgggttaag gtcgtagaag agaaggcttt ctcacccgag
      541 gttattccaa tgttctctgc cctgtccgaa ggcgctactc ctcaagacct taatacaatg
      601 ctcaataccg tgggcggtca ccaagctgct atgcagatgt tgaaggaaac tattaatgag
      661 gaagcagcag aatgggaccg agttcaccct gttcatgctg gcccaatcgc cccaggccag
      721 atgcgggaac cacggggctc tgacatagca ggaacaacat caaccttgca agagcagatc
      781 ggttggatga caaataatcc accaatacct gttggcgaaa tttacaagag gtggattata
      841 ttggggctta acaaaatcgt gcggatgtac tccccaacat caattctcga catcaggcaa
      901 ggcccaaaag aacctttccg agactatgta gaccgatttt acaaaaccct tcgagcagaa
      961 caggcttcac aagaggtaaa aaattggatg actgagacat tgttggttca gaacgccaac
     1021 cccgactgta agactattct gaaagccctg ggacccgccg caacacttga agaaatgatg
     1081 acagcctgtc agggcgtggg aggcccaggt cacaaagcac gggtgctggc tgaagcaatg
     1141 tctcaagtca ctaattctgc aactatcatg atgcagcgag ggaatttcag gaaccaacgg
     1201 aagatcgtga agtgctttaa ttgtgggaag gagggccaca ctgcccgaaa ttgtcgcgct
     1261 cccaggaaaa aaggctgctg gaagtgtggt aaagagggtc accaaatgaa agactgtact
     1321 gaaagacagg caaactttct tggtaaaata tggcctagtt acaaggggag gcccggcaat
     1381 ttcctccaga gtaggcctga gcctaccgca cccccagagg aatcattcag aagcggagtt
     1441 gagactacaa ctccccccca gaaacaagaa cctatcgata aagaactcta tcccctcacc
     1501 tcccttcgct cattgtttgg caatgaccca tctagccaat gataagaatt ctaccgggta
     1561 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     1621 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     1681 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     1741 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     1801 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     1861 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     1921 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     1981 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     2041 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     2101 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     2161 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     2221 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     2281 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     2341 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     2401 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     2461 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     2521 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     2581 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     2641 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     2701 cgaccgaaag gagcgcacga ccccatgcat cggcgtctcg agatatcagt ggtccaggct
     2761 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     2821 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     2881 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     2941 gagaagttca gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag
     3001 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     3061 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     3121 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     3181 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     3241 cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc
     3301 gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg
     3361 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3421 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3481 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg
     3541 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3601 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3661 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3721 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3781 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3841 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg
     3901 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     3961 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4021 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4081 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4141 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4201 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4261 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4321 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4381 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4441 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4501 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4561 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4621 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4681 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4741 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4801 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4861 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4921 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4981 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5041 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5101 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5161 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5221 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5281 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5341 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5401 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5461 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5521 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     5581 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5641 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5701 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5761 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5821 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5881 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5941 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag
     6001 cttggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     6061 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     6481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     6661 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcttccgata gactgcgtcg
     6721 cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggtctcgc
     6781 tgttccttgg gagggtctcc tctgagtgat tgactaccca cgacgggggt ctttcatttg
     6841 ggggctcgtc cgggatttgg agacccctgc ccagggacca ccgacccacc accgggaggt
     6901 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatgt ttgatgttat
     6961 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     7021 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     7081 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     7141 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     7201 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     7261 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     7321 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     7381 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     7441 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     7501 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     7561 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     7621 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     7681 ctccctttat ccagccctca ctccttctct aggcgccgga attgaagatc tcgatgaggc
     7741 cctttcgtct tcactcgagt ttactcccta tcagtgatag agaacgtatg aagagtttac
     7801 tccctatcag tgatagagaa cgtatgcaga ctttactccc tatcagtgat agagaacgta
     7861 taaggagttt actccctatc agtgatagag aacgtatgac cagtttactc cctatcagtg
     7921 atagagaacg tatctacagt ttactcccta tcagtgatag agaacgtata tccagtttac
     7981 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtgggc gcctataaaa
     8041 gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt
     8101 atactt
//
