LOCUS       sequence-251937-        7730 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 7730)
  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 7730)
  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
FEATURES             Location/Qualifiers
     source          1..7730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        134..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        514..717
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     668..688
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    719..737
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    740..758
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     768..792
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             892..915
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             916..942
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    958..982
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3383..3407)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3460..3501
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3581..3598)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3587..3811
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3944..3963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4095..4142
                     /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             4150..5172
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4150..5172
                     /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    5302..5423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5339..5358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5393..5412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5472..5488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5472..5488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5485..5507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5496..5512
                     /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(5520..5526,5527..5544,5545..5550))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5565..5586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5703..5720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5874..6462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5954..5973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6633..7424,7425..7493))
                     /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     7256..7275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7494..7598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7673..7692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
       61 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      121 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      541 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      601 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      661 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      721 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg
      781 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
      841 gaccgatcca gcctccggac tctagcgttt aaacttaagc ttggtaccat ggactacaag
      901 gacgacgatg acaagtaccc ttatgacgtg cccgattacg ctttgaacca gatatcaaca
      961 agtttgtaca aaaaagcagg cttcatggcc ctccacagcc cgcagtatat ttttggagat
     1021 tttagccctg atgaattcaa tcaattcttt gtgactcctc gatcttcagt tgagcttcct
     1081 ccatacagtg gaacagttct gtgtggcaca caggctgtgg ataaactacc tgatggacaa
     1141 gaatatcaga gaattgagtt tggtgtcgat gaagtcattg aacccagtga cactttgccg
     1201 agaaccccca gctacagtat ttcaagcaca ctgaaccctc aggcccctga atttattctc
     1261 ggttgtacag cttccaaaat aacccctgat ggtatcacta aagaagcaag ctatggctcc
     1321 atcgactgcc agtacccagg ctctgccctc gctttggatg gaagttctaa tgtggaggcg
     1381 gaagttttgg aaaatgatgg tgtctcaggt ggtcttggac aaagggagcg taaaaagaag
     1441 aaaaagcggc cacctggata ttacagctat ttgaaagatg gtggcgatga tagtatctcc
     1501 acagaagccc tggtcaatgg ccatgccaat tcagcagtcc cgaacagtgt cagtgcagag
     1561 gatgcagaat ttatgggtga catgcccccg tcagttacgc ccaggacttg taacagcccc
     1621 cagaactcca cagactctgt cagtgacatt gtgcctgaca gtcctttccc cggagcactc
     1681 ggcagtgaca ccaggactgc agggcagcca gaggggggcc ccggggctga ttttggtcag
     1741 tcctgcttcc ctgcagaggc tggcagagac accctgtcaa ggacagctgg ggctcagccc
     1801 tgcgttggta ccgatactac tgaaaacctt ggagttgcta atggacaaat acttgaatcc
     1861 tcgggtgagg gcacagctac caacggggtg gagttgcaca ccacggaaag catagacttg
     1921 gacccaacca aacccgagag tgcatcacct cctgctgacg gcacgggctc tgcatcaggc
     1981 acccttcctg tcagccagcc caagtcctgg gccagcctct ttcatgattc taagccctct
     2041 tcctcctcgc cggtggccta tgtggaaact aagtattccc ctcccgccat atctcccctg
     2101 gtttctgaaa agcaggttga agtcaaagaa gggcttgttc cggtttcaga ggatcctgta
     2161 gccataaaga ttgcagagtt gctggagaat gtaaccctaa tccataaacc agtgtcgttg
     2221 caaccccgtg ggctgatcaa taaagggaac tggtgctaca ttaatgctac actgcaggca
     2281 ttggttgctt gcccgccgat gtaccacctg atgaagttca ttcctctgta ttccaaagtg
     2341 caaaggcctt gtacgtcaac acccatgata gacagctttg ttcggctaat gaatgagttc
     2401 actaatatgc cagtacctcc aaaaccccga caagctcttg gagataaaat cgtgagggat
     2461 attcgccctg gagctgcctt tgagcccaca tatatttaca gactcctgac agttaacaag
     2521 tcaagcctgt ctgaaaaggg tcgacaagaa gatgctgagg aatacttagg cttcattcta
     2581 aatggacttc atgaggaaat gttgaaccta aagaagcttc tctcaccaag taatgaaaaa
     2641 cttacgattt ccaacggccc caaaaaccac tcggtcaatg aagaagagca ggaagaacaa
     2701 ggtgaaggaa gcgaggatga atgggaacaa gtgggccccc ggaacaagac ttccgtcacc
     2761 cgccaggcgg attttgttca gactccaatc accggcattt ttggtggaca catcaggtct
     2821 gtggtttacc agcagagttc aaaagaatct gccactttgc agccattttt cacgttgcag
     2881 ttggatatcc agtcagacaa gatacgcaca gtccaggatg cactggagag cttggtggca
     2941 agagaatctg tccaaggtta taccacaaaa accaaacaag aggttgagat aagtcgaaga
     3001 gtgactctgg aaaaactccc tcctgtcctc gtgctgcacc tgaaacgatt cgtttatgag
     3061 aagactggtg ggtgccagaa gcttatcaaa aatattgaat atcctgtgga cttggaaatt
     3121 agtaaagaac tgctttctcc aggggttaaa aataagaatt ttaaatgcca ccgaacctat
     3181 cggctgtttg cagtggtcta ccatcacggc aacagtgcga cgggcggcca ttacactaca
     3241 gacgtcttcc agatcggtct gaatggctgg ctgcgcatcg atgaccagac agtcaaggtg
     3301 atcaaccagt accaggtggt gaaaccaact gctgaacgca cagcctacct cctgtattac
     3361 cgccgagtgg acctgctgta atacccagct ttcttgtaca aagtggttga tatccagcac
     3421 agtggcggcc gctcaagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct
     3481 ctcctcggtc tcgattctac gcgtacctga tccgcggccg catagataac tgatccagtg
     3541 tgcctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3601 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3661 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3721 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3781 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3841 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3901 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3961 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4021 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4081 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
     4141 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
     4201 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
     4261 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
     4321 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
     4381 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
     4441 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
     4501 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
     4561 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
     4621 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
     4681 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
     4741 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
     4801 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
     4861 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
     4921 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
     4981 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
     5041 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
     5101 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
     5161 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5221 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5281 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5341 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5401 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5461 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5521 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5581 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5641 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5701 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5761 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5821 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5881 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5941 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6001 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6061 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6121 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6181 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6241 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6301 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7621 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     7681 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
//
