LOCUS       sequence-421240-        8236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Deletes PIDD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8236)
  AUTHORS   Nam M, Xia W, Mir AH, Jerrett A, Spinelli JB, Huang TT, Possemato R
  TITLE     Glucose limitation protects cancer cells from apoptosis induced by 
            pyrimidine restriction and replication inhibition.
  JOURNAL   Nat Metab. 2024 Dec;6(12):2338-2353. doi: 
            10.1038/s42255-024-01166-w. Epub 2024 Nov 26.
  PUBMED    39592843
REFERENCE   2  (bases 1 to 8236)
  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"; doi: 
            "10.1038/s42255-024-01166-w"; journalName: "Nat Metab"; date: 
            "2024-12"; volume: "6"; issue: "12"; pages: "2338-2353"
COMMENT     Sequence Label: LV-sgRNA-mCherry-Puro sgPIDD
FEATURES             Location/Qualifiers
     source          1..8236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..64)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(462..473)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(582..598)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(583..599)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1108..1288
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1350..1938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1430..1449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2109..2900,2901..2969))
                     /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     2732..2751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2970..3074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3149..3168)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3340..3719
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3721..3919
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3874..3894
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3937..4117
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4164..4289
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4786..5019
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5204..5248
                     /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"
     misc_feature    5546..5663
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5762..6012
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5762..5782
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5933..5952
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             7413..7466
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7467..8174
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(7609..7627)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7877..7896
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    join(8227..8236,1..579)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
ORIGIN
        1 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
       61 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
      121 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
      181 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
      241 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
      301 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
      361 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
      421 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
      481 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
      541 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat
      601 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
      661 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
      721 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
      781 gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
      841 taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga
      901 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa
      961 gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat
     1021 gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg
     1081 gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg
     1141 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     1201 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1261 ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg
     1321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2041 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2221 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3061 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     3121 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3181 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     3241 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     3301 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     3361 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     3421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     3481 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     3541 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     3601 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3661 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     3721 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     3781 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3841 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     3901 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
     3961 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     4021 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     4081 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     4141 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     4201 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     4261 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     4321 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     4381 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     4441 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     4501 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     4561 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     4621 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     4681 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     4741 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     4801 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     4861 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     4921 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     4981 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     5041 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     5101 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     5161 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     5221 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     5281 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     5341 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     5401 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     5461 agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca
     5521 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     5581 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     5641 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt
     5701 tagtaccggg cccgggtcga ctagtgatca gtgtgaggga gtgtaaagct ggtttcccag
     5761 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     5821 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     5881 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     5941 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     6001 accttaaaca ccggtgggcg atggctgcaa cgggtttaag agctatgctg gaaacagcat
     6061 agcaagttta aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     6121 tttttgaaac cgctgttcct aggaatcccg aggcctcgag accctgtctc acaaaataaa
     6181 gtaagcccgg actgagtgcg gaaaggcggg cctggcgggt ctggtctccc catgcgggcc
     6241 accagaggcc ctgcagcctt cagtcgcttg aaggggtaat ggcgcttcca ctcacaaaca
     6301 tggcggacag agcgtgtgaa cgagatgaac agcccctcaa aaatatggcc gccgaggctg
     6361 gacggccgtg ccccagcagc accgcctccg cgccccacgt gatctctcgc cgggcacagc
     6421 gctgaccgcg gaggtccaac cggaagaatg tccggattgg acattcggaa gagggcccgc
     6481 cttccctggg gaatctctgc gcacgcgcag aacgcttcga ccaatgaaaa cacaggaagc
     6541 cgtccgcgca accgcgttgc gtcacttctg ccgcccctgt ttcaaggtat atagccgtag
     6601 acggaacttc gcctttctct cggccttagc gccatttttt tgggtgagtg ttttttggtt
     6661 cctgcgttgg gattccgtgt acaatccata gacatctgac ctcggcactt agcatcatca
     6721 cagcaaacta actgtagcct ttctctcttt ccctgtagaa acctctgcgg atctatttcc
     6781 ggtgaattct gactgactga gccaccatga ccgagtataa gcctacagtc aggctcgcca
     6841 cacgtgatga tgtgcccagg gccgtcagaa cactggccgc tgcttttgct gactaccccg
     6901 ccaccaggca caccgtggat cccgacaggc acatcgagag agtcaccgag ctgcaggagc
     6961 tcttcctgac cagggtcggc ctggacatcg gaaaggtgtg ggtggctgac gacggcgctg
     7021 ctgtggccgt gtggacaaca cctgaaagcg tggaggccgg cgccgtgttt gccgagattg
     7081 gacccaggat ggccgaactg agcggcagca gactggctgc ccagcagcaa atggagggcc
     7141 tgctggcccc tcataggccc aaggagcctg cctggttcct ggccaccgtg ggagtgagcc
     7201 ccgatcacca gggcaaaggc ctgggcagcg ctgtggtgct gcctggagtg gaggctgccg
     7261 aaagagccgg cgtgcctgct ttcctggaga cctccgcccc cagaaacctg cccttttacg
     7321 agaggctcgg cttcaccgtg accgccgacg tggaggtccc tgaaggaccc aggacctggt
     7381 gcatgaccag aaaacccggc gctggaagcg gcgaaggaag gggctccctg ctgacatgcg
     7441 gcgatgtgga agagaaccct ggccccgtga gcaagggcga ggaggataac atggccatca
     7501 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     7561 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     7621 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     7681 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     7741 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     7801 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     7861 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     7921 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     7981 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     8041 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     8101 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     8161 agctgtacaa gtagaaacgt tgttgtttgg ggttgaatta ctcttctaga ccgcggcgcg
     8221 cccgataatc aacctc
//
