LOCUS       sequence-251548-        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_K138R
FEATURES             Location/Qualifiers
     source          1..8064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        130..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        511..709
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             727..907
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    954..1079
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1576..1809
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1994..2038
                     /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    2336..2453
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2536..2915
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2916..3119
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3145..3164
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3145..3163
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3743..3772
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3791..3814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3848..4421
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3871..4423
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4015..4032)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4242..4261
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4423..5022
                     /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(4423..4442)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4919..4939
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(5052..5085)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5141..5729
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5194..5214)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5612..5623)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5732..5748)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5733..5749)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6258..6438
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6500..7088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6580..6599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7165..7824)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7739..7758
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7825..7929)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8004..8023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
       61 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      121 tatcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
      721 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      781 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
      841 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      901 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
      961 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     1021 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     1081 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     1141 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     1201 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     1261 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     1321 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     1381 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc
     1441 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1501 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1561 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1621 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     1681 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     1741 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     1801 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     1861 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     1921 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     1981 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     2041 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     2101 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     2161 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     2221 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     2281 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     2341 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     2401 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     2461 attacaggga cagcagagat ccagtttggt tagtaccggg cccgctctag acatgtccaa
     2521 tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     2581 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     2641 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     2701 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     2761 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     2821 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     2881 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     2941 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     3001 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     3061 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     3121 gtttagtgaa ccgtcagaat tttgtaatac gactcactat agggcggccg ggaattcgtc
     3181 gactggatcc ggtaccgagg agatctgccg ccgcgatcgc cggcgcgcca tgttgatgcc
     3241 taagaagaac cggattgcca tttatgaact cctttttaag gagggagtca tggtggccag
     3301 gaaggatgtc cacatgccta agcacccgga gctggcagac aagaatgtgc ccaaccttca
     3361 tgtcatgaag gccatgcagt ctctcaagtc ccgaggctac gtgaaggaac agtttgcctg
     3421 gagacatttc tactggtacc ttaccaatga gggtatccag tatctccgtg attaccttca
     3481 tctgcccccg gagattgtgc ctgccaccct acgccgtagc cgtccagaga ctggcaggcc
     3541 tcggcctaaa ggtctggagg gtgagcgacc tgcgagactc acaagagggg aagctgacag
     3601 agatacctac agacggagtg ctgtgccacc tggtgccgac aggaaagccg aggctggggc
     3661 tgggtcagca accgaattcc agtttagagg cggatttggt cgtggacgtg gtcagccacc
     3721 tcagacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc tggcagcaaa
     3781 tgatatcctg gattacaagg atgacgacga taaggtttaa acctaggcgt agcggccgca
     3841 aattccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     3901 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     3961 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4021 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     4081 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     4141 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     4201 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     4261 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     4321 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     4381 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatgaccga gtacaagccc
     4441 acggtgcgcc tcgccacccg cgacgacgtc ccccgggcag tacgcaccct cgccgccgcg
     4501 ttcgccgact accccgccac gcgccacacc gtcgatccag accgccacat cgagcgggtc
     4561 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4621 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4681 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4741 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4801 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4861 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4921 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4981 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gatgtacaag taggattcgt
     5041 cgagggacct aataacttcg tatagcatac attatacgaa gttatacatg tttaagggtt
     5101 ccggttccac taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     5161 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5221 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5281 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5341 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5401 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5461 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5521 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     5581 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5641 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5701 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag
     5761 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc
     5821 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac
     5881 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5941 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac
     6001 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga
     6061 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg
     6121 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
     6181 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc
     6241 tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6301 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6361 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6421 tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6601 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6661 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6721 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6841 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6901 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6961 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7021 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7081 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7141 cacgttaagg gattttggtc atgattacgc cccgccctgc cactcatcgc agtactgttg
     7201 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     7261 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     7321 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     7381 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     7441 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     7501 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     7561 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaact ccggatgagc
     7621 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     7681 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     7741 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     7801 atatccagtg atttttttct ccatactctt cctttttcaa tattattgaa gcatttatca
     7861 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7921 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7981 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8041 ctgctccctg cttgtgtgtt ggag
//
