LOCUS       sequence-439197-        7263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible expression of YopE in mammalian cells by retroviral 
            transduction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7263)
  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 7263)
  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-YopE
FEATURES             Location/Qualifiers
     source          1..7263
                     /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        713..1212
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(748..771)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1145..1164
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1233..1832
                     /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(1233..1252)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1729..1749
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2013..2438
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(2541..2563)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2699..2719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2704..3033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2884..3019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3055..3074
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3155..3172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3326..3914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3406..3425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4085..4876,4877..4945))
                     /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     4708..4727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4946..5050)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5118..5136
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        5236..5615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5616..5819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5820..5995
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    6058..6257
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6458..6874
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6804..6826
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6842..6858
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        6914..7263
                     /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    6922..6940
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6958..6976
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6994..7012
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7030..7048
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7066..7084
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7102..7120
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7138..7156
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7202..7226
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ggatccgcca ccatggacta caaagacgat gacgacaaga tgaaaatcag ttctttcatt
       61 agcactagcc ttcctctgcc agcctcagtc tctgggtcaa gtagcgtagg ggaaatgtcc
      121 ggtcgatccg tgagtcagca aaagagcgat cagtacgcta acaatctggc agggcggact
      181 gagtcaccac aggggtcatc actcgccagt cggataattg aacgcctgtc tagcatggca
      241 catagtgtaa ttggtttcat acaacgaatg ttcagcgagg gtagccacaa gcctgtcgta
      301 acacctgcac tcactcccgc tcaaatgccc tctccaacat ctttctctga ttccatcaag
      361 caacttgctg ccgaaacact cccaaaatat atgcagcaac tgtcttccct tgatgctgag
      421 actctccaaa agaatcacga tcaatttgca accggttcag gccctctccg agggagtatt
      481 acccaatgtc aaggtctcat gcagttctgc gggggagaac ttcaagctga ggccagtgcc
      541 atattgaaca cccccgtatg cggaatccct ttctcccaat ggggaactgt cgggggagcc
      601 gcttccgcat acgtggcaag cggcgtcgat ctgactcagg ctgctaacga aataaagggt
      661 cttggacagc aaatgcagca gcttttgtct ctgatgtgat aagaattcta ccgggtaggg
      721 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
      781 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
      841 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
      901 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
      961 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     1021 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     1081 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     1141 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     1201 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     1261 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     1321 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     1381 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     1441 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     1501 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     1561 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     1621 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     1681 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     1741 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     1801 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     1861 ccgaaaggag cgcacgaccc catgcatcgg cgtctcgaga tatcagtggt ccaggctcta
     1921 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     1981 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     2041 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     2101 aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa ccatcagatg
     2161 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     2221 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     2281 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
     2341 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
     2401 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg
     2461 agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg cctcgcgcgt
     2521 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2581 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2641 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg
     2701 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2761 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2881 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2941 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3001 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat
     3061 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     3121 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3181 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3241 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3301 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3361 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3421 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3481 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3541 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3601 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3661 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3721 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3781 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3841 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3901 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3961 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4021 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4081 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4141 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4201 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4261 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4321 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4381 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4441 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4501 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4561 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4621 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4681 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     4741 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4801 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4861 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4921 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4981 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5041 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5101 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattagctt
     5161 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa
     5221 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     5281 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     5341 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     5401 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     5461 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     5521 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     5581 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     5641 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     5701 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5761 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     5821 aataaaagag cccacaaccc ctcactcggc gcgccagtct tccgatagac tgcgtcgccc
     5881 gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gtctcgctgt
     5941 tccttgggag ggtctcctct gagtgattga ctacccacga cgggggtctt tcatttgggg
     6001 gctcgtccgg gatttggaga cccctgccca gggaccaccg acccaccacc gggaggtaag
     6061 ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgtttg atgttatgcg
     6121 cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga
     6181 cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt
     6241 ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt
     6301 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
     6361 ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct
     6421 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
     6481 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
     6541 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
     6601 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
     6661 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
     6721 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
     6781 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
     6841 cctttatcca gccctcactc cttctctagg cgccggaatt gaagatctcg atgaggccct
     6901 ttcgtcttca ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc
     6961 ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa
     7021 ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata
     7081 gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc
     7141 ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggcgcc tataaaagca
     7201 gagctcgttt agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata
     7261 ctt
//
