LOCUS       sequence-205496-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suppression of SOD2.
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 shSOD2
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..260
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     12..32
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     183..202
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    357..474
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        523..1033
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     952..970
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1055..1654
                     /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(1055..1074)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1551..1571
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1782..2015
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2087..2208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2124..2143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2178..2197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2248..2383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2403..2422)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2404..2422)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2432..2449)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2432..2448)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2441..2463)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2590..3045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2677..2696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2887..2908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3071..3175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3176..3244,3245..4036)
                     /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(3394..3413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4207..4795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4696..4715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4949..4966
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5083..5104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5119..5124,5125..5142,5143..5149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5157..5173
                     /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     5162..5184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5181..5197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5181..5197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5216..5236
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5218..5236
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5264..5490
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5491..5671
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5718..5843
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6336..6569
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6754..6798
                     /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"
ORIGIN
        1 cccccttcac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
       61 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
      121 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      181 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      241 tgtggaaagg acgaaacacc gggtggtggt catatcaatc atactcgagt atgattgata
      301 tgaccaccac tttttgaatt ctcgacctcg agacaaatgg cagtattcat ccacaatttt
      361 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
      421 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
      481 tattacaggg acagcagaga tccactttgg ccgcggctcg agggggttgg ggttgcgcct
      541 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
      601 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
      661 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
      721 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
      781 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
      841 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
      901 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
      961 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     1021 cctctctccc cagggggatc caccggagct taccatgacc gagtacaagc ccacggtgcg
     1081 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     1141 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     1201 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     1261 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     1321 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     1381 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     1441 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     1501 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     1561 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     1621 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     1681 gaccgaaagg agcgcacgac cccatgcatc ggtaccttta agaccaatga cttacaaggc
     1741 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     1801 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     1861 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     1921 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     1981 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     2041 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     2101 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2161 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     2221 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2281 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2341 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     2401 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     2461 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2521 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2581 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2641 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2701 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2761 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2821 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2881 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2941 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3001 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     3061 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3121 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3181 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3241 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3301 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3361 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3421 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3481 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3541 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3601 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3661 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3721 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3781 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3841 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3901 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3961 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4021 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4081 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4141 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4201 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4261 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4321 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4381 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4441 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4501 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4561 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4621 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4681 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4741 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4801 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4861 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4921 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4981 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     5041 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     5101 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     5161 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     5221 taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata
     5281 ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa
     5341 aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg
     5401 caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt
     5461 gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc
     5521 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5581 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5641 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
     5701 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     5761 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     5821 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     5881 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     5941 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     6001 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     6061 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     6121 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     6181 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     6241 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     6301 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     6361 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     6421 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     6481 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     6541 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     6601 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     6661 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     6721 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     6781 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     6841 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     6901 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     6961 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     7021 agatccattc gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg
     7081 gccg
//
