LOCUS       sequence-251584-        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
FEATURES             Location/Qualifiers
     source          1..8298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    74..191
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        274..653
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        654..857
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     883..902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        883..901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1715..1744
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1763..1786
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1820..2393
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1843..2395
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1987..2004)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2214..2233
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2395..2994
                     /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(2395..2414)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2891..2911
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(3024..3057)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3113..3701
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3166..3186)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3584..3595)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3704..3720)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3705..3721)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4230..4410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4472..5060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4552..4571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5137..5796)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5711..5730
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5797..5901)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5976..5995)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6166..6545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6547..6745
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6763..6943
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6990..7115
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7612..7845
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8030..8074
                     /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 atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag
       61 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
      121 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
      181 ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgggcc
      241 cgctctagac atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      301 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      361 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      421 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      481 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      541 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      601 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      661 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      721 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      781 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      841 tctatataag cagagctcgt ttagtgaacc gtcagaattt tgtaatacga ctcactatag
      901 ggcggccggg aattcgtcga ctggatccgg taccgaggag atctgccgcc gcgatcgccg
      961 gcgcgccatg gcagtgcaaa tatccaagaa gaggaagttt gtcgctgatg gcatcttcaa
     1021 agctgaactg aatgagtttc ttactcggga gctggctgaa gatggctact ctggagttga
     1081 ggtgcgagtt acaccaacca ggacagaaat cattatctta gccaccagaa cacagaatgt
     1141 tcttggtgag aagggccggc ggattcggga actgactgct gtagttcaga agaggtttgg
     1201 ctttccagag ggcagtgtag agctttatgc tgaaaaggtg gccactagag gtctgtgtgc
     1261 cattgcccag gcagagtctc tgcgttacaa actcctagga gggcttgctg tgcggagggc
     1321 ctgctatggt gtgctgcggt tcatcatgga gagtggggcc aaaggctgcg aggttgtggt
     1381 gtctgggaaa ctccgaggac agagggctaa atccatgaag tttgtggatg gcctgatgat
     1441 ccacagcgga gaccctgtta actactacgt tgacactgct gtgcgccacg tgttgctcag
     1501 acagggtgtg ctgggcatca aggtgaagat catgctgccc tgggacccaa ctggtaagat
     1561 tggccctaag aagcccctgc ctgaccacgt gagcattgtg gaacccaaag atgagatact
     1621 gcccaccacc cccatctcag aacagaaggg tgggaagcca gagccgcctg ccatgcccca
     1681 gccagtcccc acagcaacgc gtacgcggcc gctcgagcag aaactcatct cagaagagga
     1741 tctggcagca aatgatatcc tggattacaa ggatgacgac gataaggttt aaacctaggc
     1801 gtagcggccg caaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     1861 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     1921 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     1981 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     2041 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     2101 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     2161 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2221 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     2281 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     2341 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatgacc
     2401 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc agtacgcacc
     2461 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc agaccgccac
     2521 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     2581 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     2641 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     2701 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     2761 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     2821 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     2881 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     2941 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatgtaca
     3001 agtaggattc gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca
     3061 tgtttaaggg ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc
     3121 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3181 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3241 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3301 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3361 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3421 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3481 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3541 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3601 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3661 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     3721 atcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     3781 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     3841 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     3901 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     3961 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     4021 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     4081 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     4141 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     4201 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     4261 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4321 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     4381 ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca
     4441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4981 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5101 ggaacgaaaa ctcacgttaa gggattttgg tcatgattac gccccgccct gccactcatc
     5161 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     5221 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     5281 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     5341 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     5401 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     5461 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     5521 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     5581 ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     5641 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     5701 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     5761 atcaacggtg gtatatccag tgattttttt ctccatactc ttcctttttc aatattattg
     5821 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5881 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     5941 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6001 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6061 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     6121 aggcgttttg cgctgcttcg cgatgtacgg gccagatatc gcgttgacat tgattattga
     6181 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6241 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6301 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6361 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6421 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6481 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6541 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6601 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6661 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6721 tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac
     6781 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6841 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6901 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     6961 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     7021 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     7081 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     7141 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     7201 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     7261 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     7321 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     7381 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     7441 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     7501 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     7561 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     7621 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     7681 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     7741 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     7801 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     7861 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     7921 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     7981 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     8041 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     8101 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     8161 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     8221 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     8281 agagagagac agagacag
//
