LOCUS       sequence-198941-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suppression of FXN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  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 7083)
  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 shFXN_1
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..517
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     436..454
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             539..1138
                     /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(539..558)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1035..1055
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1266..1499
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1571..1692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1608..1627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1662..1681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1732..1867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1887..1906)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1888..1906)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1916..1933)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1916..1932)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1925..1947)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2074..2529
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2161..2180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2371..2392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2555..2659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2660..2728,2729..3520)
                     /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(2878..2897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3691..4279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4180..4199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4433..4450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4567..4588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4603..4608,4609..4626,4627..4633)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4641..4657
                     /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     4646..4668
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4665..4681
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4665..4681
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4700..4720
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4702..4720
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4748..4974
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4975..5155
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5202..5327
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5820..6053
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6238..6282
                     /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        6580..6828
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6580..6600
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6751..6770
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6924..7041
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
       61 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
      121 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
      181 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
      241 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
      301 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
      361 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
      421 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
      481 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat
      541 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
      601 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
      661 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
      721 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
      781 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
      841 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
      901 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
      961 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     1021 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     1081 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     1141 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc
     1201 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     1261 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     1321 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     1381 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     1441 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     1501 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     1561 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1681 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     1741 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     1801 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     1861 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     1921 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1981 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2041 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2101 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2161 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2221 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2281 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2341 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2401 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2461 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     2521 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2581 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2641 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2701 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2761 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2821 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2881 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2941 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3001 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3061 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3121 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3181 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3241 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3301 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3361 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3421 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3481 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3541 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3601 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4261 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4321 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4381 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4441 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4501 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4561 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4621 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4681 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     4741 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     4801 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     4861 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     4921 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     4981 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5041 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5101 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     5161 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     5221 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     5281 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     5341 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     5401 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     5461 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     5521 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     5581 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     5641 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     5701 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     5761 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     5821 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     5881 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     5941 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     6001 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     6061 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     6121 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     6181 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     6241 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     6301 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     6361 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     6421 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     6481 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     6541 cgatcacgag actagcctcg agcggccgcc cccttcaccg agggcctatt tcccatgatt
     6601 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
     6661 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     6721 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     6781 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg gctggactct
     6841 ttagcagagt tctcgagaac tctgctaaag agtccagctt tttaattctc gacctcgaga
     6901 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     6961 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     7021 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggccg
     7081 cgg
//
