LOCUS       sequence-358365-        6322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP tagged PKM2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6322)
  AUTHORS   Stetak A, Veress R, Ovadi J, Csermely P, Keri G, Ullrich A
  TITLE     Nuclear translocation of the tumor marker pyruvate kinase M2 induces
            programmed cell death.
  JOURNAL   Cancer Res. 2007 Feb 15;67(4):1602-8.
  PUBMED    17308100
REFERENCE   2  (bases 1 to 6322)
  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"; journalName: "Cancer 
            Res."; date: "2007-02-15"; volume: "67(4)"; pages: "1602-8"
COMMENT     Sequence Label: pEGFP-C1-PKM2
FEATURES             Location/Qualifiers
     source          1..6322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      548..557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(554..556,557..559,560..1270)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(599..620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(860..879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1207..1228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3051..3172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3088..3107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3142..3161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3179..3634)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3316..3337)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3528..3547
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3661..3765
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3767..4124
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3767..3958
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3790..3810)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3975..4110
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4159..4953
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4159..4953
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4213..4232)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4823..4842
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5141..5160)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5185..5232
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5561..6149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6050..6069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1141 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1261 gctgtacaag tccggactca gatccacctc agcagccatg tcgaagcccc atagtgaagc
     1321 cgggactgcc ttcattcaga cccagcagct gcacgcagcc atggctgaca cattcctgga
     1381 gcacatgtgc cgcctggaca ttgattcacc acccatcaca gcccggaaca ctggcatcat
     1441 ctgtaccatt ggcccagctt cccgatcagt ggagacgttg aaggagatga ttaagtctgg
     1501 aatgaatgtg gctcgtctga acttctctca tggaactcat gagtaccatg cggagaccat
     1561 caagaatgtg cgcacagcca cggaaagctt tgcttctgac cccatcctct accggcccgt
     1621 tgctgtggct ctagacacta aaggacctga gatccgaact gggctcatca agggcagcgg
     1681 cactgcagag gtggagctga agaagggagc cactctcaaa atcacgctgg ataacgccta
     1741 catggaaaag tgtgacgaga acatcctgtg gctggactac aagaacatct gcaaggtggt
     1801 ggaagtgggc agcaagatct acgtggatga tgggcttatt tctctccagg tgaagcagaa
     1861 aggtgccgac ttcctggtga cggaggtgga aaatggtggc tccttgggca gcaagaaggg
     1921 tgtgaacctt cctggggctg ctgtggactt gcctgctgtg tcggagaagg acatccagga
     1981 tctgaagttt ggggtcgagc aggatgttga tatggtgttt gcgtcattca tccgcaaggc
     2041 atctgatgtc catgaagtta ggaaggtcct gggagagaag ggaaagaaca tcaagattat
     2101 cagcaaaatc gagaatcatg agggggttcg gaggtttgat gaaatcctgg aggccagtga
     2161 tgggatcatg gtggctcgtg gtgatctagg cattgagatt cctgcagaga aggtcttcct
     2221 tgctcagaag atgatgattg gacggtgcaa ccgagctggg aagcctgtca tctgtgctac
     2281 tcagatgctg gagagcatga tcaagaagcc ccgccccact cgggctgaag gcagtgatgt
     2341 ggccaatgca gtcctggatg gagccgactg catcatgctg tctggagaaa cagccaaagg
     2401 ggactatcct ctggaggctg tgcgcatgca gcacctgatt gcccgtgagg cagaggctgc
     2461 catctaccac ttgcaattat ttgaggaact ccgccgcctg gcgcccatta ccagcgaccc
     2521 cacagaagcc accgccgtgg gtgccgtgga ggcctccttc aagtgctgca gtggggccat
     2581 aatcgtcctc accaagtctg gcaggtctgc tcaccaggtg gccagatacc gcccacgtgc
     2641 ccccatcatt gctgtgaccc ggaatcccca gacagctcgt caggcccacc tgtaccgtgg
     2701 catcttccct gtgctgtgca aggacccagt ccaggaggcc tgggctgagg acgtggacct
     2761 ccgggtgaac tttgccatga atgttggcaa ggcccgaggc ttcttcaaga agggagatgt
     2821 ggtcattgtg ctgaccggat ggcgccctgg ctccggcttc accaacacca tgcgtgttgt
     2881 tcctgtgccg gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat
     2941 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3001 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3061 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3121 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     3181 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3241 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3301 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3361 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3421 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3481 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3541 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3601 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3661 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3721 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3781 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3841 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3901 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3961 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4021 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4081 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4141 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4201 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4261 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4321 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4381 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4441 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4501 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4561 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4621 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4681 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4741 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4801 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4861 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4921 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4981 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5041 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5101 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5161 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5221 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5281 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5341 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5401 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5461 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5521 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5581 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5641 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5701 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5761 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5821 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5881 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5941 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6001 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6061 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6121 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6181 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6241 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     6301 catagcccat atatggagtt cc
//
