LOCUS       sequence-332992-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suppress ACO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Terzi EM, Sviderskiy VO, Alvarez SW, Whiten GC, Possemato R
  TITLE     Iron-sulfur cluster deficiency can be sensed by IRP2 and regulates 
            iron homeostasis and sensitivity to ferroptosis independent of IRP1 
            and FBXL5.
  JOURNAL   Sci Adv. 2021 May 26;7(22). pii: 7/22/eabg4302. doi: 
            10.1126/sciadv.abg4302. Print 2021 May.
  PUBMED    34039609
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: "Sci Adv."; 
            date: "2021-05-26"; volume: "7(22). pii: 7/22/eabg4302. doi"; pages:
            " 10.1126/sciadv.abg4302. Print 2021 May"
COMMENT     Sequence Label: pLKO.1P shACO1
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..519
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     438..456
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             541..1140
                     /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(541..560)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1037..1057
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1268..1501
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1573..1694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1610..1629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1664..1683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1734..1869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1889..1908)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1890..1908)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1918..1935)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1918..1934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1927..1949)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2076..2531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2163..2182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2373..2394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2557..2661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2662..2730,2731..3522)
                     /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(2880..2899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3693..4281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4182..4201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4435..4452
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4569..4590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4605..4610,4611..4628,4629..4635)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4643..4659
                     /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     4648..4670
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4667..4683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4667..4683
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4702..4722
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4704..4722
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4750..4976
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4977..5157
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5204..5329
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5822..6055
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6240..6284
                     /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        6582..6830
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6582..6602
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6753..6772
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6927..7044
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctcgaggg ggttggggtt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc
       61 ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc
      121 ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg
      181 acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg
      241 tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca
      301 atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc
      361 gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc
      421 ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc
      481 ggctccctcg ttgaccgaat caccgacctc tctccccagg gggatccacc ggagcttacc
      541 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
      601 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
      661 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
      721 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
      781 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
      841 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
      901 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
      961 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     1021 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     1081 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     1141 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcggta
     1201 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     1261 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     1321 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     1381 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     1441 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     1501 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     1561 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1621 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1681 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     1741 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1801 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     1861 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     1921 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1981 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2041 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     2101 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2161 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2221 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2281 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2341 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2401 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2461 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2521 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2581 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2641 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2701 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2761 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2821 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2881 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2941 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3001 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3061 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3121 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3181 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3241 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3301 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3361 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3421 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3481 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3541 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3601 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3661 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3721 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3781 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3841 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3901 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3961 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4021 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4081 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4141 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4201 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4261 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4321 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4381 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4441 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4501 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4561 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4621 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4681 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     4741 caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
     4801 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
     4861 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
     4921 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
     4981 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5041 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5101 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     5161 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     5221 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     5281 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     5341 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     5401 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     5461 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     5521 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     5581 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     5641 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     5701 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     5761 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     5821 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     5881 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     5941 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     6001 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     6061 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     6121 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     6181 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     6241 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     6301 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     6361 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     6421 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     6481 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
     6541 atcgatcacg agactagcct cgagcggccg cccccttcac cgagggccta tttcccatga
     6601 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga
     6661 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     6721 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     6781 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggccaggaaa
     6841 gaaattcttc aatctcgaga ttgaagaatt tctttcctgg tttttgaatt ctcgacctcg
     6901 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     6961 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     7021 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     7081 ccgc
//
