LOCUS       sequence-251649-        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_K138/139R
FEATURES             Location/Qualifiers
     source          1..8064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        163..542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        543..746
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..791
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        772..790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1370..1399
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1418..1441
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1475..2048
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1498..2050
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1642..1659)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1869..1888
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2050..2649
                     /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(2050..2069)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2546..2566
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(2679..2712)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2768..3356
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2821..2841)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3239..3250)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3359..3375)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3360..3376)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3885..4065
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4127..4715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4207..4226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4792..5451)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5366..5385
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5452..5556)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5631..5650)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5821..6200
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6202..6400
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6418..6598
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6645..6770
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7267..7500
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7685..7729
                     /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    join(8027..8064,1..80)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
       61 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
      121 taccgggccc gctctagaca tgtccaatat gaccgccatg ttgacattga ttattgacta
      181 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      241 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      301 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      361 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      421 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      481 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      541 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      601 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      661 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      721 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      781 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
      841 cgatcgccgg cgcgccatgt tgatgcctaa gaagaaccgg attgccattt atgaactcct
      901 ttttaaggag ggagtcatgg tggccaggaa ggatgtccac atgcctaagc acccggagct
      961 ggcagacaag aatgtgccca accttcatgt catgaaggcc atgcagtctc tcaagtcccg
     1021 aggctacgtg aaggaacagt ttgcctggag acatttctac tggtacctta ccaatgaggg
     1081 tatccagtat ctccgtgatt accttcatct gcccccggag attgtgcctg ccaccctacg
     1141 ccgtagccgt ccagagactg gcaggcctcg gcctaaaggt ctggagggtg agcgacctgc
     1201 gagactcaca agaggggaag ctgacagaga tacctacaga cggagtgctg tgccacctgg
     1261 tgccgacagg agagccgagg ctggggctgg gtcagcaacc gaattccagt ttagaggcgg
     1321 atttggtcgt ggacgtggtc agccacctca gacgcgtacg cggccgctcg agcagaaact
     1381 catctcagaa gaggatctgg cagcaaatga tatcctggat tacaaggatg acgacgataa
     1441 ggtttaaacc taggcgtagc ggccgcaaat tccgcccctc tccctccccc ccccctaacg
     1501 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1561 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     1621 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1681 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1741 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1801 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1861 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1921 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     1981 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2041 gatgataata tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     2101 cgggcagtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     2161 gatccagacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     2221 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     2281 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     2341 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     2401 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     2461 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     2521 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     2581 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     2641 ggtgcctgat gtacaagtag gattcgtcga gggacctaat aacttcgtat agcatacatt
     2701 atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     2761 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2821 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2881 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2941 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3001 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3061 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3121 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3181 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3241 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3301 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3361 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     3421 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     3481 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     3541 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     3601 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     3661 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     3721 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     3781 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     3841 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     3901 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     3961 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     4021 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4561 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4741 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg attacgcccc
     4801 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     4861 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     4921 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     4981 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     5041 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     5101 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     5161 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     5221 cattgccata cggaactccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     5281 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     5341 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     5401 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca tactcttcct
     5461 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5521 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5581 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5641 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5701 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     5761 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatcgcgtt
     5821 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     5881 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     5941 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6001 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6061 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6121 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6181 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     6241 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     6301 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     6361 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
     6421 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6481 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6541 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6601 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     6661 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     6721 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     6781 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     6841 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     6901 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     6961 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     7021 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     7081 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     7141 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     7201 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     7261 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     7321 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     7381 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     7441 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     7501 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     7561 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     7621 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     7681 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     7741 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     7801 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     7861 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     7921 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     7981 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     8041 gggggattgg ggggtacagt gcag
//
