LOCUS       sequence-439196-        7326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible expression of SopE in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7326)
  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 7326)
  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-SopE
FEATURES             Location/Qualifiers
     source          1..7326
                     /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        776..1275
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(811..834)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1208..1227
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1296..1895
                     /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(1296..1315)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1792..1812
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2076..2501
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(2604..2626)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2762..2782)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2767..3096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2947..3082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3118..3137
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3218..3235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3389..3977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3469..3488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4148..4939,4940..5008))
                     /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     4771..4790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5009..5113)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5181..5199
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        5299..5678
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5679..5882
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5883..6058
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    6121..6320
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6521..6937
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6867..6889
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6905..6921
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        6977..7326
                     /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    6985..7003
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7021..7039
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7057..7075
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7093..7111
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7129..7147
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7165..7183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7201..7219
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7265..7289
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ggatccgcca ccatggacta caaagacgat gacgacaaga tgactaaaat tacactgtct
       61 cctcagaact ttcgaatcca gaaacaagag actaccctgc ttaaagagaa gtcaaccgaa
      121 aagaactccc ttgccaagag catcttggca gtaaagaatc actttataga gctgaggtcc
      181 aaactgtctg aaagatttat tagccacaaa aacacagaat ccagcgctac ccattttcat
      241 agaggctcag catctgaagg acgagctgta cttaccaata aagtggtcaa ggattttatg
      301 ctccagactc tgaatgacat agacatccga gggagtgctt ctaaggaccc tgcttacgca
      361 tcccaaacac gggaagccat attgagcgca gtctattcaa agaacaagga ccaatgctgc
      421 aatcttctta ttagcaaggg gatcaatata gccccattcc tgcaagaaat tggagaggcc
      481 gcaaagaacg ccgggcttcc aggcaccacc aagaacgacg tgttcacacc atcaggagct
      541 ggggctaacc cttttattac accactcatt agttcagcta actccaagta cccacgaatg
      601 ttcattaacc aacatcagca agcctcattt aagatttacg cagaaaagat catcatgacc
      661 gaagtggctc ccctcttcaa tgaatgtgca atgccaaccc ctcagcagtt ccagctcatt
      721 cttgagaata tagctaacaa atatattcag aacacccctt gataagaatt ctaccgggta
      781 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
      841 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
      901 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
      961 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     1021 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     1081 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     1141 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     1201 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     1261 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     1321 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     1381 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     1441 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     1501 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     1561 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     1621 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     1681 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     1741 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     1801 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     1861 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     1921 cgaccgaaag gagcgcacga ccccatgcat cggcgtctcg agatatcagt ggtccaggct
     1981 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     2041 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     2101 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     2161 gagaagttca gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag
     2221 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     2281 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     2341 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     2401 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     2461 cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc
     2521 gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg
     2581 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     2641 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     2701 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg
     2761 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2821 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2941 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3001 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3061 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg
     3121 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     3181 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3241 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3301 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4441 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4741 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     4801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5161 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag
     5221 cttggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     5281 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     5341 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     5401 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5461 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5521 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5581 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5641 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     5701 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     5761 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5821 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5881 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcttccgata gactgcgtcg
     5941 cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggtctcgc
     6001 tgttccttgg gagggtctcc tctgagtgat tgactaccca cgacgggggt ctttcatttg
     6061 ggggctcgtc cgggatttgg agacccctgc ccagggacca ccgacccacc accgggaggt
     6121 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatgt ttgatgttat
     6181 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     6241 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     6301 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     6361 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     6421 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     6481 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     6541 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     6601 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     6661 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     6721 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     6781 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     6841 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     6901 ctccctttat ccagccctca ctccttctct aggcgccgga attgaagatc tcgatgaggc
     6961 cctttcgtct tcactcgagt ttactcccta tcagtgatag agaacgtatg aagagtttac
     7021 tccctatcag tgatagagaa cgtatgcaga ctttactccc tatcagtgat agagaacgta
     7081 taaggagttt actccctatc agtgatagag aacgtatgac cagtttactc cctatcagtg
     7141 atagagaacg tatctacagt ttactcccta tcagtgatag agaacgtata tccagtttac
     7201 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtgggc gcctataaaa
     7261 gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt
     7321 atactt
//
