LOCUS       sequence-251553-        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_K139R
FEATURES             Location/Qualifiers
     source          1..8064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        321..524
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..569
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        550..568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1148..1177
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1196..1219
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1253..1826
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1276..1828
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1420..1437)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1647..1666
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1828..2427
                     /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(1828..1847)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2324..2344
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(2457..2490)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2546..3134
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2599..2619)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3017..3028)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3137..3153)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3138..3154)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3663..3843
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3905..4493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3985..4004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4570..5229)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5144..5163
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5230..5334)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5409..5428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5599..5978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5980..6178
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6196..6376
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6423..6548
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7045..7278
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7463..7507
                     /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    7805..7922
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        join(8005..8064,1..320)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
       61 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      121 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      181 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      241 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      301 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      361 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      421 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      481 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      541 agaattttgt aatacgactc actatagggc ggccgggaat tcgtcgactg gatccggtac
      601 cgaggagatc tgccgccgcg atcgccggcg cgccatgttg atgcctaaga agaaccggat
      661 tgccatttat gaactccttt ttaaggaggg agtcatggtg gccaggaagg atgtccacat
      721 gcctaagcac ccggagctgg cagacaagaa tgtgcccaac cttcatgtca tgaaggccat
      781 gcagtctctc aagtcccgag gctacgtgaa ggaacagttt gcctggagac atttctactg
      841 gtaccttacc aatgagggta tccagtatct ccgtgattac cttcatctgc ccccggagat
      901 tgtgcctgcc accctacgcc gtagccgtcc agagactggc aggcctcggc ctaaaggtct
      961 ggagggtgag cgacctgcga gactcacaag aggggaagct gacagagata cctacagacg
     1021 gagtgctgtg ccacctggtg ccgacaagag agccgaggct ggggctgggt cagcaaccga
     1081 attccagttt agaggcggat ttggtcgtgg acgtggtcag ccacctcaga cgcgtacgcg
     1141 gccgctcgag cagaaactca tctcagaaga ggatctggca gcaaatgata tcctggatta
     1201 caaggatgac gacgataagg tttaaaccta ggcgtagcgg ccgcaaattc cgcccctctc
     1261 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     1321 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     1381 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     1441 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     1501 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     1561 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     1621 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     1681 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     1741 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     1801 ttttcctttg aaaaacacga tgataatatg accgagtaca agcccacggt gcgcctcgcc
     1861 acccgcgacg acgtcccccg ggcagtacgc accctcgccg ccgcgttcgc cgactacccc
     1921 gccacgcgcc acaccgtcga tccagaccgc cacatcgagc gggtcaccga gctgcaagaa
     1981 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     2041 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     2101 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     2161 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     2221 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     2281 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     2341 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     2401 tgcatgaccc gcaagcccgg tgcctgatgt acaagtagga ttcgtcgagg gacctaataa
     2461 cttcgtatag catacattat acgaagttat acatgtttaa gggttccggt tccactaggt
     2521 acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2581 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2641 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2701 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2761 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2821 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2881 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2941 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3001 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3061 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3121 ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa
     3181 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     3241 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     3301 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     3361 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     3421 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     3481 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     3541 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     3601 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     3661 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     3721 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     3781 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     3841 gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4321 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4561 tggtcatgat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     4621 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     4681 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     4741 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     4801 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     4861 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     4921 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     4981 caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc
     5041 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     5101 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     5161 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     5221 tttctccata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5281 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5341 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     5401 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     5461 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     5521 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     5581 cgggccagat atcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     5641 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     5701 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5761 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5821 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5881 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5941 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     6001 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6061 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6121 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg
     6181 cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6241 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6301 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6361 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     6421 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     6481 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     6541 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     6601 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     6661 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     6721 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     6781 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     6841 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     6901 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     6961 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     7021 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     7081 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     7141 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     7201 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     7261 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     7321 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     7381 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     7441 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     7501 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     7561 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     7621 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     7681 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     7741 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     7801 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     7861 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     7921 ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga
     7981 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     8041 ttacggggtc attagttcat agcc
//
