LOCUS       sequence-430272-        8484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8484)
  AUTHORS   Anastasiou D, Poulogiannis G, Asara JM, Boxer MB, Jiang JK, Shen M, 
            Bellinger G, Sasaki AT, Locasale JW, Auld DS, Thomas CJ, Vander 
            Heiden MG, Cantley LC
  TITLE     Inhibition of pyruvate kinase M2 by reactive oxygen species 
            contributes to cellular antioxidant responses.
  JOURNAL   Science. 2011 Dec 2;334(6060):1278-83. doi: 10.1126/science.1211485.
            Epub 2011 Nov 3.
  PUBMED    22052977
REFERENCE   2  (bases 1 to 8484)
  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: 
            "10.1126/science.1211485"; journalName: "Science"; date: "2011-12-2-
            2"; volume: "334"; issue: "6060"; pages: "1278-83"
COMMENT     Sequence Label: pLHCX-Flag-mPKM2
FEATURES             Location/Qualifiers
     source          1..8484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      690..699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             699..722
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             2406..2999
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3102..3124)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3210..3350
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3265..3284
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3365..3382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3536..4124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3616..3635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4295..5086,5087..5155))
                     /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     4918..4937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5156..5260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5328..5346
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5918..6093
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    6156..6355
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6556..6972
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6902..6924
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6940..6956
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             7028..8047
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             7028..8047
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccccaag cttgttaacg ccaccatgga ctacaaagac gatgacgata
      721 aagcaaggga tccactagta acggccgcca gtgtgctgga attcatgccg aagccacaca
      781 gtgaagcagg gactgccttc attcagaccc agcagctcca tgcagccatg gctgacacct
      841 tcctggaaca catgtgccgc ctggacattg actctgcccc catcacggcc cgcaacactg
      901 gcatcatttg taccattggg cctgcttccc gatctgtgga gatgctgaag gagatgatta
      961 agtctggaat gaatgtggct cggctgaatt tctctcatgg aacccatgag taccatgcag
     1021 agaccatcaa gaatgtccgt gaagccacag aaagctttgc atctgatccc attctctacc
     1081 gtcctgttgc ggtggctctg gatacaaagg gacctgagat ccggactgga ctcatcaagg
     1141 gcagcggcac cgctgaggtg gagctgaaga agggagccac tctgaagatc accctggaca
     1201 acgcttacat ggagaagtgt gacgagaaca tcctgtggct ggactacaag aacatctgca
     1261 aggtggtgga ggtgggcagc aagatctacg tggacgatgg gctcatctca ctgcaggtga
     1321 aggagaaagg cgctgacttc ctggtgacgg aggtggagaa tggtggctcc ttgggcagca
     1381 agaagggcgt gaacctgccg ggcgctgctg tggatctccc cgctgtgtcg gaaaaggaca
     1441 tccaggacct gaagtttggg gtggagcagg atgtggacat ggtgtttgca tctttcatcc
     1501 gcaaggcagc cgacgtgcat gaagtcagga aggtgctggg agagaagggc aagaacatca
     1561 agatcatcag caaaatcgag aaccatgaag gcgtccgcag gtttgatgag atcttggagg
     1621 ccagtgatgg gatcatggtg gctcgtggtg acctgggcat tgagattcct gcagagaagg
     1681 tcttcctggc tcagaagatg atgatcgggc gatgcaaccg agctgggaag cctgtcatct
     1741 gtgccacaca gatgctggag agcatgatca agaagccacg ccccacccgt gctgaaggca
     1801 gtgatgtggc caatgcagtc ctggatggag cagactgcat catgctgtct ggagaaacag
     1861 ccaaggggga ctaccctctg gaggctgttc gcatgcagca cctgattgcc cgagaggcag
     1921 aggctgccat ctaccacttg cagctattcg aggaactccg ccgcctggcg cccattacca
     1981 gcgaccccac agaagctgcc gccgtgggtg ccgtggaggc ctccttcaag tgctgcagtg
     2041 gggccattat cgtgctcacc aagtctggca ggagtgctca ccaagtggcc aggtaccgcc
     2101 ctcgggctcc tatcattgcc gtgactcgaa atccccagac tgctcgccag gcccatctgt
     2161 accgtggcat cttccctgtg ctgtgtaagg atgccgtgct gaatgcctgg gctgaggatg
     2221 tcgaccttcg tgtaaacttg gccatggatg ttggcaaggc ccgaggcttc ttcaagaagg
     2281 gagatgtggt cattgtgctg accgggtggc gccctggctc tggattcacc aacaccatgc
     2341 gtgtagtgcc tgtaccttga atcgataaaa taaaagattt tatttagtct ccagaaaaag
     2401 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     2461 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
     2521 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     2581 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     2641 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     2701 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     2761 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     2821 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
     2881 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     2941 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
     3001 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
     3061 taagctggct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3121 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3181 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     3241 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3301 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     3361 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3421 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3481 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3541 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3601 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3661 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3721 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3781 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3841 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3901 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3961 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4021 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4081 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4141 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4201 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4261 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4321 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4381 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4441 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4501 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4561 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     4621 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4681 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4741 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4801 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4861 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     4921 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4981 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5041 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5101 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5161 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5221 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5281 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5341 cgaggccctt tcgtcttcaa gaattcatac cagatcaccg aaaactgtcc tccaaatgtg
     5401 tccccctcac actcccaaat tcgcgggctt ctgcctctta gaccactcta ccctattccc
     5461 cacactcacc ggagccaaag ccgcggccct tccgtttctt tgcttttgaa agaccccacc
     5521 cgtaggtggc aagctagctt aagtaacgcc actttgcaag gcatggaaaa atacataact
     5581 gagaatagaa aagttcagat caaggtcagg aacaaagaaa cagctgaata ccaaacagga
     5641 tatctgtggt aagcggttcc tgccccggct cagggccaag aacagatgag acagctgagt
     5701 gatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcggg gccaagaaca
     5761 gatggtcccc agatgcggtc cagccctcag cagtttctag tgaatcatca gatgtttcca
     5821 gggtgcccca aggacctgaa aatgaccctg taccttattt gaactaacca atcagttcgc
     5881 ttctcgcttc tgttcgcgcg cttccgctct ccgagctcaa taaaagagcc cacaacccct
     5941 cactcggcgc gccagtcttc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa
     6001 gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg tctcctctga
     6061 gtgattgact acccacgacg ggggtctttc atttgggggc tcgtccggga tttggagacc
     6121 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt
     6181 ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc tgcgtctgta ctagttagct
     6241 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
     6301 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
     6361 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
     6421 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
     6481 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     6541 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     6601 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     6661 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     6721 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     6781 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     6841 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     6901 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     6961 tctctaggcg ccggaattcc gatctgatag cttgccacaa cccgtaccaa agatggatag
     7021 atccggaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     7081 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     7141 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     7201 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     7261 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     7321 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     7381 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     7441 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     7501 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     7561 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     7621 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     7681 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     7741 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     7801 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     7861 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     7921 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     7981 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     8041 ggaatagagt agatgccgac cgaacaagag ctgatttcga gaacgcctca gccagcaact
     8101 cgcgcgagcc tagcaaggca aatgcgagag aacggcctta cgcttggtgg cacagttctc
     8161 gtccacagtt cgctaagctc gctcggctgg gtcgcgggag ggccggtcgc agtgattcag
     8221 gcccttctgg attgtgttgg tccccagggc acgattgtca tgcccacgca ctcgggtgat
     8281 ctgactgatc ccgcagattg gagatcgccg cccgtgcctg ccgattgggt gcagatctct
     8341 agagctcgct gatccggcca ttagccatat tattcattgg ttatatagca taaatcaata
     8401 ttggctattg gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
     8461 catgtccaac attaccgcca tgtt
//
