LOCUS       sequence-194971-        7466 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Suppression of ISCU.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7466)
  AUTHORS   Alvarez SW, Sviderskiy VO, Terzi EM, Papagiannakopoulos T, Moreira 
            AL, Adams S, Sabatini DM, Birsoy K, Possemato R
  TITLE     NFS1 undergoes positive selection in lung tumours and protects cells
            from ferroptosis.
  JOURNAL   Nature. 2017 Nov 22. doi: 10.1038/nature24637.
  PUBMED    29168506
REFERENCE   2  (bases 1 to 7466)
  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: "Nature. 
            2017 Nov 22. doi: 10.1038/nature24637."
COMMENT     Sequence Label: pLKO.1P shISCU
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    42..159
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        208..718
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     637..655
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             740..1339
                     /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(740..759)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1236..1256
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1355..1943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1408..1428)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1826..1837)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2015..2248
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2326..2447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2363..2382)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2417..2436
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2487..2622
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2642..2661)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2643..2661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2671..2688)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2671..2687)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2680..2702)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2829..3284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2916..2935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3126..3147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3310..3414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3415..3483,3484..4275)
                     /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(3633..3652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4446..5034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4935..4954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5188..5205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5322..5343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5358..5363,5364..5381,5382..5388)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5396..5412
                     /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     5401..5423
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5420..5436
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5420..5436
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5455..5475
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5457..5475
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5503..5729
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5730..5910
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5957..6082
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6575..6808
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6993..7037
                     /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        7215..7463
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7215..7235
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7386..7405
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
       61 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
      121 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
      181 agagatccac tttggcgccg gctcgagggg gcccgggttg cgccttttcc aaggcagccc
      241 tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca gcggcgccga
      301 ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct tcgccgctac
      361 ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag gttccttgcg
      421 gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta ccctcgcaga
      481 cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg ccaatagcgg
      541 ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg aggcggggtg
      601 tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct gcaagcctcc
      661 ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct ctccccaggg
      721 ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
      781 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
      841 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
      901 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgccgtggc
      961 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     1021 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     1081 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     1141 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     1201 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     1261 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     1321 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     1381 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1441 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1501 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1561 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1621 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1681 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     1741 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     1801 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     1861 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     1921 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     1981 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     2041 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     2101 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     2161 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     2221 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     2281 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     2341 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2401 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     2461 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2521 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2581 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     2641 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     2701 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     2761 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     2821 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2881 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2941 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3001 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3061 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3121 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3181 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3241 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     3301 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3361 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3421 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3481 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3541 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3601 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3661 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3721 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3781 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3841 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3901 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3961 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4021 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4081 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4141 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4201 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4261 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4321 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4381 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5101 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5161 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5221 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5281 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     5341 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     5401 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     5461 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     5521 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     5581 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     5641 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     5701 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     5761 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     5821 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     5881 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     5941 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     6001 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     6061 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     6121 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     6181 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     6241 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     6301 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     6361 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     6421 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     6481 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     6541 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     6601 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     6661 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     6721 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     6781 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     6841 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     6901 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     6961 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     7021 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     7081 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     7141 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     7201 cgaggggacc cagagagggc ctatttccca tgattccttc atatttgcat atacgataca
     7261 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     7321 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     7381 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     7441 tcttgtggaa aggacgaggt accggt
//
