LOCUS       sequence-243134-        7771 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 7771)
  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 7771)
  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: pFRT_TO_FlagHA_USP10_C424A
FEATURES             Location/Qualifiers
     source          1..7771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..42
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(86..105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        277..656
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        657..860
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     811..831
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    862..880
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    883..901
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     911..935
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1035..1058
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1059..1085
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1101..1125
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3525..3549)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3602..3643
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3723..3740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3729..3953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4086..4105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4237..4284
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4292..5314
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4292..5314
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5444..5565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5481..5500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5535..5554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5614..5630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5614..5630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5627..5649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5638..5654
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5662..5668,5669..5686,5687..5692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5707..5728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5845..5862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6016..6604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6096..6115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6775..7566,7567..7635))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7398..7417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7636..7740)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cggacggatc gggagatctc
       61 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      121 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
      181 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
      241 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
      301 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      361 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      421 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      481 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      541 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      601 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      661 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      721 gtctccaccc cattgacgtc aatgggagtt tgttttggaa ccaaaatcaa cgggactttc
      781 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      841 aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc agtgatagag
      901 atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
      961 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta
     1021 agcttggtac catggactac aaggacgacg atgacaagta cccttatgac gtgcccgatt
     1081 acgctttgaa ccagatatca acaagtttgt acaaaaaagc aggcttcatg gccctccaca
     1141 gcccgcagta tatttttgga gattttagcc ctgatgaatt caatcaattc tttgtgactc
     1201 ctcgatcttc agttgagctt cctccataca gtggaacagt tctgtgtggc acacaggctg
     1261 tggataaact acctgatgga caagaatatc agagaattga gtttggtgtc gatgaagtca
     1321 ttgaacccag tgacactttg ccgagaaccc ccagctacag tatttcaagc acactgaacc
     1381 ctcaggcccc tgaatttatt ctcggttgta cagcttccaa aataacccct gatggtatca
     1441 ctaaagaagc aagctatggc tccatcgact gccagtaccc aggctctgcc ctcgctttgg
     1501 atggaagttc taatgtggag gcggaagttt tggaaaatga tggtgtctca ggtggtcttg
     1561 gacaaaggga gcgtaaaaag aagaaaaagc ggccacctgg atattacagc tatttgaaag
     1621 atggtggcga tgatagtatc tccacagaag ccctggtcaa tggccatgcc aattcagcag
     1681 tcccgaacag tgtcagtgca gaggatgcag aatttatggg tgacatgccc ccgtcagtta
     1741 cgcccaggac ttgtaacagc ccccagaact ccacagactc tgtcagtgac attgtgcctg
     1801 acagtccttt ccccggagca ctcggcagtg acaccaggac tgcagggcag ccagaggggg
     1861 gccccggggc tgattttggt cagtcctgct tccctgcaga ggctggcaga gacaccctgt
     1921 caaggacagc tggggctcag ccctgcgttg gtaccgatac tactgaaaac cttggagttg
     1981 ctaatggaca aatacttgaa tcctcgggtg agggcacagc taccaacggg gtggagttgc
     2041 acaccacgga aagcatagac ttggacccaa ccaaacccga gagtgcatca cctcctgctg
     2101 acggcacggg ctctgcatca ggcacccttc ctgtcagcca gcccaagtcc tgggccagcc
     2161 tctttcatga ttctaagccc tcttcctcct cgccggtggc ctatgtggaa actaagtatt
     2221 cccctcccgc catatctccc ctggtttctg aaaagcaggt tgaagtcaaa gaagggcttg
     2281 ttccggtttc agaggatcct gtagccataa agattgcaga gttgctggag aatgtaaccc
     2341 taatccataa accagtgtcg ttgcaacccc gtgggctgat caataaaggg aactgggcct
     2401 acattaatgc tacactgcag gcattggttg cttgcccgcc gatgtaccac ctgatgaagt
     2461 tcattcctct gtattccaaa gtgcaaaggc cttgtacgtc aacacccatg atagacagct
     2521 ttgttcggct aatgaatgag ttcactaata tgccagtacc tccaaaaccc cgacaagctc
     2581 ttggagataa aatcgtgagg gatattcgcc ctggagctgc ctttgagccc acatatattt
     2641 acagactcct gacagttaac aagtcaagcc tgtctgaaaa gggtcgacaa gaagatgctg
     2701 aggaatactt aggcttcatt ctaaatggac ttcatgagga aatgttgaac ctaaagaagc
     2761 ttctctcacc aagtaatgaa aaacttacga tttccaacgg ccccaaaaac cactcggtca
     2821 atgaagaaga gcaggaagaa caaggtgaag gaagcgagga tgaatgggaa caagtgggcc
     2881 cccggaacaa gacttccgtc acccgccagg cggattttgt tcagactcca atcaccggca
     2941 tttttggtgg acacatcagg tctgtggttt accagcagag ttcaaaagaa tctgccactt
     3001 tgcagccatt tttcacgttg cagttggata tccagtcaga caagatacgc acagtccagg
     3061 atgcactgga gagcttggtg gcaagagaat ctgtccaagg ttataccaca aaaaccaaac
     3121 aagaggttga gataagtcga agagtgactc tggaaaaact ccctcctgtc ctcgtgctgc
     3181 acctgaaacg attcgtttat gagaagactg gtgggtgcca gaagcttatc aaaaatattg
     3241 aatatcctgt ggacttggaa attagtaaag aactgctttc tccaggggtt aaaaataaga
     3301 attttaaatg ccaccgaacc tatcggctct ttgcagtggt ctaccatcac ggcaacagtg
     3361 cgacgggcgg ccattacact acagacgtct tccagatcgg tctgaatggc tggctgcgca
     3421 tcgatgacca gacagtcaag gtgatcaacc agtaccaggt ggtgaaacca actgctgaac
     3481 gcacagccta cctcctgtat taccgccgag tggacctgct gtaaacccag ctttcttgta
     3541 caaagtggtt gatatccagc acagtggcgg ccgctcaagt ctagagggcc cgcggttcga
     3601 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gatccgcggc
     3661 cgcatagata actgatccag tgtgcctcga gtctagaggg cccgtttaaa cccgctgatc
     3721 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3781 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3841 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3901 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3961 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4201 caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta
     4261 ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt
     4321 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     4381 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     4441 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     4501 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     4561 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4621 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     4681 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4741 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4801 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4861 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     4921 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     4981 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     5041 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     5101 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     5161 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     5221 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     5281 acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac
     5341 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5401 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5461 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5521 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5581 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5641 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5701 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5761 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5821 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5881 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5941 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6001 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6061 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6121 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6421 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7621 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7741 cacatttccc cgaaaagtgc cacctgacgt c
//
