LOCUS       sequence-192206-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockdown of MCT4 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Lim KS, Lim KJ, Price AC, Orr BA, Eberhart CG, Bar EE
  TITLE     Inhibition of monocarboxylate transporter-4 depletes stem-like 
            glioblastoma cells and inhibits HIF transcriptional response in a 
            lactate-independent manner.
  JOURNAL   Oncogene. 2014 Aug 28;33(35):4433-41. doi: 10.1038/onc.2013.390. 
            Epub 2013 Sep 30.
  PUBMED    24077291
REFERENCE   2  (bases 1 to 7084)
  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: 
            "10.1038/onc.2013.390"; journalName: "Oncogene"; date: "2014-08-28- 
            28"; volume: "33"; issue: "35"; pages: "4433-41"
COMMENT     Sequence Label: plko.1/shMCT4 477
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             186..366
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    413..538
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1031..1264
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1449..1493
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        1791..2039
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1791..1811
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1962..1981
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2136..2253
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2302..2812
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2731..2749
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2834..3433
                     /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(2834..2853)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3330..3350
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3561..3794
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3866..3987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3903..3922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3957..3976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4027..4162
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4182..4201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4183..4201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4211..4228)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4211..4227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4220..4242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4369..4824
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4456..4475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4666..4687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4850..4954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4955..5023,5024..5815)
                     /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     complement(5173..5192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5986..6574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6475..6494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6728..6745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6862..6883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6898..6903,6904..6921,6922..6928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6936..6952
                     /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)."
     primer_bind     6941..6963
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6960..6976
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6960..6976
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6995..7015
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6997..7015
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(7043..7084,1..185)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
       61 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
      121 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
      181 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
      241 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      301 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      361 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
      421 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
      481 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
      541 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
      601 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
      661 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
      721 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
      781 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
      841 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
      901 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
      961 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     1021 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     1081 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     1141 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     1201 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     1261 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     1321 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     1381 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     1441 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     1501 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     1561 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     1621 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     1681 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     1741 ctcgacggta tcgatcacga gactagcctc gagcggccgc ccccttcacc gagggcctat
     1801 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     1861 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     1921 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     1981 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     2041 ggctcatcat gctgaaccgc tactcgagta gcggttcagc atgatgagct ttttgaattc
     2101 tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     2161 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2221 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     2281 ccactttggc cgcggctcga gggggttggg gttgcgcctt ttccaaggca gccctgggtt
     2341 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
     2401 gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
     2461 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
     2521 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
     2581 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
     2641 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
     2701 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
     2761 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggatcc
     2821 accggagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     2881 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     2941 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     3001 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     3061 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     3121 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     3181 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     3241 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     3301 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     3361 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     3421 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     3481 ccatgcatcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     3541 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     3601 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     3661 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     3721 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     3781 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     3841 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     3901 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3961 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     4021 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4081 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4141 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     4201 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4261 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4321 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     4381 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4441 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4501 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4561 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4621 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4681 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4741 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4801 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4921 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4981 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5041 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5101 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5161 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5221 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5281 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5341 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5401 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5461 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5521 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5581 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5641 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5701 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5761 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5821 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5881 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5941 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6001 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6061 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6121 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6601 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6661 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6721 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6781 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6841 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6901 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6961 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     7021 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
     7081 gtaa
//
