LOCUS       sequence-198952-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suppression of SOD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  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 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"; journalName: "Nature. 
            2017 Nov 22. doi: 10.1038/nature24637."
COMMENT     Sequence Label: pLKO.1P shSOD1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..521
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     440..458
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             543..1142
                     /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(543..562)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1039..1059
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1270..1503
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1575..1696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1612..1631)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1666..1685
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1736..1871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1891..1910)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1892..1910)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1920..1937)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1920..1936)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1929..1951)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2078..2533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2375..2396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2559..2663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2664..2732,2733..3524)
                     /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(2882..2901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3695..4283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4184..4203
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4437..4454
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4571..4592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4607..4612,4613..4630,4631..4637)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4645..4661
                     /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     4650..4672
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4669..4685
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4669..4685
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4704..4724
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4706..4724
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4752..4978
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4979..5159
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5206..5331
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5824..6057
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6242..6286
                     /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        6584..6832
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6584..6604
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6755..6774
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6929..7046
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gcggctcgag ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac
       61 gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat
      121 tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg
      181 cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga
      241 cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag
      301 caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg
      361 ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg
      421 ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag
      481 tcggctccct cgttgaccga atcaccgacc tctctcccca gggggatcca ccggagctta
      541 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
      601 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
      661 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
      721 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
      781 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
      841 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
      901 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
      961 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     1021 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     1081 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     1141 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg
     1201 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     1261 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     1321 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     1381 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     1441 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1501 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     1561 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1621 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1681 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     1741 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     1801 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     1861 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     1921 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     1981 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2041 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     2101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2221 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2281 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2341 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2401 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2461 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2521 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2581 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2641 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2701 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2761 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2821 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2881 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2941 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3001 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3061 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3121 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3181 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3241 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3301 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3361 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3421 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3481 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3541 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3601 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3661 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3721 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3781 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3841 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3901 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3961 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4021 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4081 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4141 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4201 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4261 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4321 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4381 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4441 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     4501 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     4561 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     4621 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     4681 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     4741 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
     4801 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
     4861 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
     4921 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
     4981 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     5041 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     5101 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     5161 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     5221 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     5281 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     5341 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     5401 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     5461 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     5521 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     5581 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     5641 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     5701 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     5761 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     5821 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     5881 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     5941 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     6001 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     6061 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     6121 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     6181 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     6241 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     6301 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     6361 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     6421 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     6481 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
     6541 gtatcgatca cgagactagc ctcgagcggc cgcccccttc accgagggcc tatttcccat
     6601 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     6661 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     6721 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     6781 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggagcaga
     6841 tgacttgggc aaaggctcga gcctttgccc aagtcatctg cttttttgaa ttctcgacct
     6901 cgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     6961 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     7021 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt
     7081 ggcc
//
