LOCUS       sequence-251583-        8447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8447)
  AUTHORS   Meyer C, Garzia A, Morozov P, Molina H, Tuschl T
  TITLE     The G3BP1-Family-USP10 Deubiquitinase Complex Rescues Ubiquitinated 
            40S Subunits of Ribosomes Stalled in Translation from Lysosomal 
            Degradation.
  JOURNAL   Mol Cell. 2020 Mar 19;77(6):1193-1205.e5. doi: 
            10.1016/j.molcel.2019.12.024. Epub 2020 Jan 24.
  PUBMED    31981475
REFERENCE   2  (bases 1 to 8447)
  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: 
            "10.1016/j.molcel.2019.12.024"; journalName: "Mol Cell"; date: 
            "2020-03-19- 19"; volume: "77"; issue: "6"; pages: "1193-1205.e5"
COMMENT     Sequence Label: pLENTI_RPS2
FEATURES             Location/Qualifiers
     source          1..8447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    156..273
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        356..735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        736..939
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     965..984
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        965..983
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1946..1975
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1994..2017
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2051..2624
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2074..2626
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2218..2235)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2445..2464
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2626..3225
                     /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(2626..2645)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3122..3142
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(3255..3288)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3344..3932
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3397..3417)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3815..3826)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3935..3951)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3936..3952)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4461..4641
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4703..5291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4783..4802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5368..6027)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5942..5961
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6028..6132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6207..6226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6397..6776
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6778..6976
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6994..7174
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7221..7346
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7843..8076
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8261..8305
                     /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 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
       61 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
      121 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
      181 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
      241 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
      301 ccagtttggt tagtaccggg cccgctctag acatgtccaa tatgaccgcc atgttgacat
      361 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      421 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      481 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      541 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      601 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      661 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
      721 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
      781 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      841 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
      901 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagaat
      961 tttgtaatac gactcactat agggcggccg ggaattcgtc gactggatcc ggtaccgagg
     1021 agatctgccg ccgcgatcgc cggcgcgtat ggcggatgac gccggtgcag cgggggggcc
     1081 cgggggccct ggtggccctg ggatggggaa ccgcggtggc ttccgcggag gtttcggcag
     1141 tggcatccgg ggccggggtc gcggccgtgg acggggccgg ggccgaggcc gcggagctcg
     1201 cggaggcaag gccgaggata aggagtggat gcccgtcacc aagttgggcc gcttggtcaa
     1261 ggacatgaag atcaagtccc tggaggagat ctatctcttc tccctgccca ttaaggaatc
     1321 agagatcatt gatttcttcc tgggggcctc tctcaaggat gaggttttga agattatgcc
     1381 agtgcagaag cagacccgtg ccggccagcg caccaggttc aaggcatttg ttgctatcgg
     1441 ggactacaat ggccacgtcg gtctgggtgt taagtgctcc aaggaggtgg ccaccgccat
     1501 ccgtggggcc atcatcctgg ccaagctctc catcgtcccc gtgcgcagag gctactgggg
     1561 gaacaagatc ggcaagcccc acactgtccc ttgcaaggtg acaggccgct gcggctctgt
     1621 gctggtacgc ctcatccctg cacccagggg cactggcatc gtctccgcac ctgtgcctaa
     1681 gaagctgctc atgatggctg gtatcgatga ctgctacacc tcagcccggg gctgcactgc
     1741 caccctgggc aacttcgcca aggccacctt tgatgccatt tctaagacct acagctacct
     1801 gacccccgac ctctggaagg agactgtatt caccaagtct ccctatcagg agttcactga
     1861 ccacctcgtc aagacccaca ccagagtctc cgtgcagcgg actcaggctc cagctgtggc
     1921 tacaacaacg cgtacgcggc cgctcgagca gaaactcatc tcagaagagg atctggcagc
     1981 aaatgatatc ctggattaca aggatgacga cgataaggtt taaacctagg cgtagcggcc
     2041 gcaaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     2101 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     2161 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     2221 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     2281 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     2341 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     2401 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2461 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2521 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2581 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatgac cgagtacaag
     2641 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg cagtacgcac cctcgccgcc
     2701 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cagaccgcca catcgagcgg
     2761 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2821 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     2881 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     2941 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3001 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3061 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3121 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     3181 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgatgtac aagtaggatt
     3241 cgtcgaggga cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg
     3301 gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta
     3361 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3421 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3481 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3541 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3601 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3661 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3721 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     3781 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3841 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3901 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc
     3961 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     4021 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     4081 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     4141 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     4201 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
     4261 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
     4321 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     4381 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
     4441 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     4501 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4561 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4621 cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5101 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5341 actcacgtta agggattttg gtcatgatta cgccccgccc tgccactcat cgcagtactg
     5401 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     5461 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     5521 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     5581 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     5641 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     5701 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     5761 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga actccggatg
     5821 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     5881 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     5941 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     6001 ggtatatcca gtgatttttt tctccatact cttccttttt caatattatt gaagcattta
     6061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6121 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6181 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6241 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     6301 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     6361 gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca ttgattattg actagttatt
     6421 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     6481 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     6541 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     6601 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     6661 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     6721 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     6781 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6841 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6901 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     6961 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
     7021 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7081 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7141 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     7201 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     7261 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     7321 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     7381 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     7441 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     7501 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     7561 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     7621 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     7681 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7741 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7801 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7861 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     7921 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     7981 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8041 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8101 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8161 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8221 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8281 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8341 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8401 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgt
//
