LOCUS       sequence-251586-        8064 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 8064)
  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 8064)
  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_RPS10
FEATURES             Location/Qualifiers
     source          1..8064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    292..409
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        492..871
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        872..1075
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1101..1120
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1101..1119
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1699..1728
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1747..1770
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1804..2377
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1827..2379
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1971..1988)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2198..2217
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2379..2978
                     /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(2379..2398)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2875..2895
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(3008..3041)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3097..3685
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3150..3170)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3568..3579)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3688..3704)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3689..3705)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4214..4394
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4456..5044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4536..4555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5121..5780)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5695..5714
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5781..5885)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5960..5979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6150..6529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6531..6729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6747..6927
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6974..7099
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7596..7829
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8014..8058
                     /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 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
       61 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
      121 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
      181 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
      241 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
      301 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
      361 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
      421 cagggacagc agagatccag tttggttagt accgggcccg ctctagacat gtccaatatg
      481 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      541 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      601 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      661 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      721 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
      781 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
      841 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
      901 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      961 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     1021 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     1081 agtgaaccgt cagaattttg taatacgact cactataggg cggccgggaa ttcgtcgact
     1141 ggatccggta ccgaggagat ctgccgccgc gatcgccggc gcgccatgtt gatgcctaag
     1201 aagaaccgga ttgccattta tgaactcctt tttaaggagg gagtcatggt ggccaggaag
     1261 gatgtccaca tgcctaagca cccggagctg gcagacaaga atgtgcccaa ccttcatgtc
     1321 atgaaggcca tgcagtctct caagtcccga ggctacgtga aggaacagtt tgcctggaga
     1381 catttctact ggtaccttac caatgagggt atccagtatc tccgtgatta ccttcatctg
     1441 cccccggaga ttgtgcctgc caccctacgc cgtagccgtc cagagactgg caggcctcgg
     1501 cctaaaggtc tggagggtga gcgacctgcg agactcacaa gaggggaagc tgacagagat
     1561 acctacagac ggagtgctgt gccacctggt gccgacaaga aagccgaggc tggggctggg
     1621 tcagcaaccg aattccagtt tagaggcgga tttggtcgtg gacgtggtca gccacctcag
     1681 acgcgtacgc ggccgctcga gcagaaactc atctcagaag aggatctggc agcaaatgat
     1741 atcctggatt acaaggatga cgacgataag gtttaaacct aggcgtagcg gccgcaaatt
     1801 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     1861 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     1921 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     1981 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     2041 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     2101 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     2161 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     2221 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     2281 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     2341 cggggacgtg gttttccttt gaaaaacacg atgataatat gaccgagtac aagcccacgg
     2401 tgcgcctcgc cacccgcgac gacgtccccc gggcagtacg caccctcgcc gccgcgttcg
     2461 ccgactaccc cgccacgcgc cacaccgtcg atccagaccg ccacatcgag cgggtcaccg
     2521 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     2581 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     2641 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     2701 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     2761 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     2821 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     2881 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     2941 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatg tacaagtagg attcgtcgag
     3001 ggacctaata acttcgtata gcatacatta tacgaagtta tacatgttta agggttccgg
     3061 ttccactagg tacaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     3121 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3181 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3241 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3301 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3361 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3421 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3481 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3541 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3601 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3661 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa
     3721 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     3781 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     3841 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     3901 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     3961 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     4021 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     4081 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     4141 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     4201 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4261 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4321 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4381 gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4861 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5101 ttaagggatt ttggtcatga ttacgccccg ccctgccact catcgcagta ctgttgtaat
     5161 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     5221 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     5281 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     5341 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     5401 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     5461 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     5521 ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg atgagcattc
     5581 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     5641 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     5701 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     5761 ccagtgattt ttttctccat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5821 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5881 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     5941 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     6001 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     6061 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     6121 ttcgcgatgt acgggccaga tatcgcgttg acattgatta ttgactagtt attaatagta
     6181 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6241 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6301 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6421 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6481 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     6541 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6601 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6661 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6721 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6781 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6841 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6901 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     6961 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     7021 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     7081 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     7141 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     7201 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     7261 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     7321 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     7381 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     7441 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     7501 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     7561 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     7621 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     7681 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     7741 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     7801 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     7861 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     7921 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     7981 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     8041 gataaatggg caagtttgtg gaat
//
