LOCUS       sequence-379334-        7985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of tamoxifen-inducible Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7985)
  AUTHORS   Shi DD, Savani MR, Levitt MM, Wang AC, Endress JE, Bird CE, Buehler 
            J, Stopka SA, Regan MS, Lin YF, Puliyappadamba VT, Gao W, Khanal J, 
            Evans L, Lee JH, Guo L, Xiao Y, Xu M, Huang B, Jennings RB, Bonal 
            DM, Martin-Sandoval MS, Dang T, Gattie LC, Cameron AB, Lee S, Asara 
            JM, Kornblum HI, Mak TW, Looper RE, Nguyen QD, Signoretti S, Gradl 
            S, Sutter A, Jeffers M, Janzer A, Lehrman MA, Zacharias LG, Mathews 
            TP, Losman JA, Richardson TE, Cahill DP, DeBerardinis RJ, Ligon KL, 
            Xu L, Ly P, Agar NYR, Abdullah KG, Harris IS, Kaelin WG Jr, McBrayer
            SK
  TITLE     De novo pyrimidine synthesis is a targetable vulnerability in IDH 
            mutant glioma.
  JOURNAL   Cancer Cell. 2022 Sep 12;40(9):939-956.e16. doi: 
            10.1016/j.ccell.2022.07.011. Epub 2022 Aug 18.
  PUBMED    35985343
REFERENCE   2  (bases 1 to 7985)
  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.ccell.2022.07.011"; journalName: "Cancer Cell"; date: 
            "2022-09-12- 12"; volume: "40"; issue: "9"; pages: "939-956.e16"
COMMENT     Sequence Label: pMSCV-MerCreMer-hygro
FEATURES             Location/Qualifiers
     source          1..7985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     222..241
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             307..1332
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             307..1332
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             1690..2204
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    2366..2382
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2390..2396,2397..2414,2415..2420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2435..2456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2573..2590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2744..3332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2824..2843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3503..4294,4295..4363))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4126..4145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4364..4468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4536..4554
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4592..4614)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4714..4733
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4927..4949
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5077..5096
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5296..5812
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5876..6217
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6284..6700
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6630..6652
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6668..6684
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        join(7775..7985,1..289)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7810..7833)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
       61 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
      121 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
      181 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
      241 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcatccc
      301 gccaccatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
      361 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
      421 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
      481 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
      541 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
      601 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
      661 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
      721 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
      781 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
      841 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
      901 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
      961 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     1021 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     1081 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     1141 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     1201 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     1261 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     1321 gcaaaggaat agagtagatg ccgaccgaac aagagctgat ttcgagaacg cctcagccag
     1381 caactcgcgc gagcctagca aggcaaatgc gagagaacgg ccttacgctt ggtggcacag
     1441 ttctcgtcca cagttcgcta agctcgctcg gctgggtcgc gggagggccg gtcgcagtga
     1501 ttcaggccct tctggattgt gttggtcccc agggcacgat tgtcatgccc acgcactcgg
     1561 gtgatctgac tgatcccgca gattggagat cgccgcccgt gcctgccgat tgggtgcaga
     1621 tccgtcgacc tgcagccaag cttatcgata aaataaaaga ttttatttag tctccagaaa
     1681 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
     1741 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
     1801 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     1861 agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca
     1921 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     1981 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     2041 cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt
     2101 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     2161 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg
     2221 aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta
     2281 gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca
     2341 gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag
     2401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2521 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3181 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3361 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3781 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3841 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3901 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3961 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4021 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4081 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4141 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4201 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4261 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4321 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4381 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4441 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4501 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4561 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4621 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4681 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4741 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     4801 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4861 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     4921 ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat
     4981 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     5041 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     5101 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt
     5161 agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata
     5221 tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct
     5281 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     5341 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     5401 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5461 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     5521 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     5581 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     5641 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     5701 tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac tcgctgatcc
     5761 ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca tttggaggtt
     5821 ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg gaggtaagct
     5881 ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg gcatctaatg
     5941 tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg
     6001 aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg
     6061 ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata
     6121 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
     6181 ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg
     6241 ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac
     6301 tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc
     6361 ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa
     6421 cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa
     6481 ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga
     6541 agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc
     6601 tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg
     6661 atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat ctgcggccgc
     6721 aagcttgcat gcctgcacac catgggagat ccacgaaatg aaatgggtgc ttcaggagac
     6781 atgagggctg ccaacctttg gccaagccct cttgtgatta agcacactaa gaagaatagc
     6841 cctgccttgt ccttgacagc tgaccagatg gtcagtgcct tgttggatgc tgaaccgccc
     6901 atgatctatt ctgaatatga tccttctaga cccttcagtg aagcctcaat gatgggctta
     6961 ttgaccaacc tagcagatag ggagctggtt catatgatca actgggcaaa gagagtgcca
     7021 ggctttgggg acttgaatct ccatgatcag gtccaccttc tcgagtgtgc ctggctggag
     7081 attctgatga ttggtctcgt ctggcgctcc atggaacacc cggggaagct cctgtttgct
     7141 cctaacttgc tcctggacag gaatcaaggt aaatgtgtgg aaggcatggt ggagatcttt
     7201 gacatgttgc ttgctacgtc aagtcggttc cgcatgatga acctgcaggg tgaagagttt
     7261 gtgtgcctca aatccatcat tttgcttaat tccggagtgt acacgtttct gtccagcacc
     7321 ttgaagtctc tggaagagaa ggaccacatc caccgtgtcc tggacaagat cacagacact
     7381 ttgatccacc tgatggccaa agctggcctg actctgcagc agcagcatcg ccgcctagct
     7441 cagctccttc tcattctttc ccatatccgg catatgagta acaaacgcat ggagcatctc
     7501 tacaacatga aatgcaaaaa cgtggtaccc ctctatgacc tgctcctgga gatgttggat
     7561 gcccaccgcc ttcatgcccc agccagtcgc atgggagtgc ccccagagga gcccagccag
     7621 acccagctgg ccaccaccag ctccacttca gcacattcct tacaaaccta ctacataccc
     7681 ccggaagcag agggcttccc caacacgatc tgagagctcc tactagtgaa ttaaacttaa
     7741 ttaaggccgg ccctcgaggt taacgaattc taccgggtag gggaggcgct tttcccaagg
     7801 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     7861 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     7921 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     7981 gcgtc
//
