LOCUS       sequence-251589-        8298 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 8298)
  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 8298)
  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_RPS3_K214R
FEATURES             Location/Qualifiers
     source          1..8298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    245..362
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        445..824
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        825..1028
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1054..1073
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1054..1072
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1886..1915
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1934..1957
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1991..2564
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2014..2566
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2158..2175)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2385..2404
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2566..3165
                     /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(2566..2585)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3062..3082
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(3195..3228)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3284..3872
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3337..3357)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3755..3766)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3875..3891)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3876..3892)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4401..4581
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4643..5231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4723..4742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5308..5967)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5882..5901
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5968..6072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6147..6166)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6337..6716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6718..6916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6934..7114
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7161..7286
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7783..8016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8201..8245
                     /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 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
       61 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
      121 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
      181 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
      241 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
      301 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
      361 ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga
      421 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
      481 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      541 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      601 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      661 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      721 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
      781 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      841 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      901 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      961 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     1021 gcagagctcg tttagtgaac cgtcagaatt ttgtaatacg actcactata gggcggccgg
     1081 gaattcgtcg actggatccg gtaccgagga gatctgccgc cgcgatcgcc ggcgcgccat
     1141 ggcagtgcaa atatccaaga agaggaagtt tgtcgctgat ggcatcttca aagctgaact
     1201 gaatgagttt cttactcggg agctggctga agatggctac tctggagttg aggtgcgagt
     1261 tacaccaacc aggacagaaa tcattatctt agccaccaga acacagaatg ttcttggtga
     1321 gaagggccgg cggattcggg aactgactgc tgtagttcag aagaggtttg gctttccaga
     1381 gggcagtgta gagctttatg ctgaaaaggt ggccactaga ggtctgtgtg ccattgccca
     1441 ggcagagtct ctgcgttaca aactcctagg agggcttgct gtgcggaggg cctgctatgg
     1501 tgtgctgcgg ttcatcatgg agagtggggc caaaggctgc gaggttgtgg tgtctgggaa
     1561 actccgagga cagagggcta aatccatgaa gtttgtggat ggcctgatga tccacagcgg
     1621 agaccctgtt aactactacg ttgacactgc tgtgcgccac gtgttgctca gacagggtgt
     1681 gctgggcatc aaggtgaaga tcatgctgcc ctgggaccca actggtaaga ttggccctaa
     1741 gaagcccctg cctgaccacg tgagcattgt ggaacccaga gatgagatac tgcccaccac
     1801 ccccatctca gaacagaagg gtgggaagcc agagccgcct gccatgcccc agccagtccc
     1861 cacagcaacg cgtacgcggc cgctcgagca gaaactcatc tcagaagagg atctggcagc
     1921 aaatgatatc ctggattaca aggatgacga cgataaggtt taaacctagg cgtagcggcc
     1981 gcaaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     2041 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     2101 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     2161 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     2221 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     2281 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     2341 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2401 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2461 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2521 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatgac cgagtacaag
     2581 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg cagtacgcac cctcgccgcc
     2641 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cagaccgcca catcgagcgg
     2701 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2761 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     2821 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     2881 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     2941 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3001 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3061 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     3121 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgatgtac aagtaggatt
     3181 cgtcgaggga cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg
     3241 gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta
     3301 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3361 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3421 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3481 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3541 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3601 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3661 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     3721 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3781 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3841 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc
     3901 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     3961 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     4021 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     4081 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     4141 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
     4201 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
     4261 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     4321 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
     4381 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     4441 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4501 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4561 cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgatta cgccccgccc tgccactcat cgcagtactg
     5341 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     5401 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     5461 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     5521 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     5581 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     5641 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     5701 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga actccggatg
     5761 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     5821 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     5881 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     5941 ggtatatcca gtgatttttt tctccatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6121 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6181 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     6241 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     6301 gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca ttgattattg actagttatt
     6361 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     6421 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     6481 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     6541 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     6601 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     6661 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     6721 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6781 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6841 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     6901 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
     6961 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7021 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7081 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     7141 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     7201 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     7261 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     7321 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     7381 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     7441 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     7501 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     7561 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     7621 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7681 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7741 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7801 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     7861 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     7921 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     7981 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8041 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8101 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8161 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8221 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8281 tataaaatta ttcataat
//
