LOCUS       sequence-152197-        8839 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 8839)
  AUTHORS   Sowa ME, Bennett EJ, Gygi SP, Harper JW
  TITLE     Defining the human deubiquitinating enzyme interaction landscape.
  JOURNAL   Cell. 2009 Jul 23. 138(2):389-403.
  PUBMED    19615732
REFERENCE   2  (bases 1 to 8839)
  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"; journalName: "Cell. 2009 
            Jul 23. 138(2):389-403."
COMMENT     Sequence Label: Flag-HA-USP10
FEATURES             Location/Qualifiers
     source          1..8839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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)."
     LTR             307..821
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(921..1520)
                     /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(1004..1024)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1501..1520
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1541..2040)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1589..1608)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1982..2005
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(2053..2079)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(2053..2079)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2095..2118)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    2150..2174
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4566..4590)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4619..4638)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4620..4638)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(4650..5066)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4666..4682)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4698..4720)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5133..5474
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             5538..6054
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6254..6273)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6401..6423)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6617..6636)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6736..6758
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6796..6814)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6882..6986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6987..7055,7056..7847)
                     /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     complement(7205..7224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8018..8606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8507..8526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8760..8777
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      181 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      241 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      301 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      361 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      421 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      481 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      541 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      601 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      661 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
      721 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
      781 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
      841 ctaaataaaa tcttttattt tatcgatgca tggggtcgtg cgctcctttc ggtcgggcgc
      901 tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
      961 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     1021 ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     1081 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     1141 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     1201 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     1261 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     1321 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     1381 gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg
     1441 aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc
     1501 gtgggcttgt actcggtcat ggtaagcttg ggctgcaggt cgaaaggccc ggagatgagg
     1561 aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg
     1621 aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac
     1681 cccttcccag cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc
     1741 aaaggcctac ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac
     1801 gtgctacttc catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc
     1861 tgactagggg aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc
     1921 gcctaccggt ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc
     1981 ccagcggggc tgctaaagcg catgctccag actgccttgg gaaaagcgcc tcccctaccc
     2041 ggtagaattc taggcgtagt cgggcacgtc gtaggggtat cctccagcgg ccgacttgtc
     2101 atcgtcgtcc ttgtagtcct cgagttcgaa ccgcgggccc tctagatcaa ccactttgta
     2161 caagaaagtt ggcagcaggt ccactcggcg gtaatacagg aggtaggctg tgcgttcagc
     2221 agttggtttc accacctggt actggttgat caccttgact gtctggtcat cgatgcgcag
     2281 ccagccattc agaccgatct ggaagacgtc tgtagtgtaa tggccgcccg tcgcactgtt
     2341 gccgtgatgg tagaccactg caaagagccg ataggttcgg tggcatttaa aattcttatt
     2401 tttaacccct ggagaaagca gttctttact aatttccaag tccacaggat attcaatatt
     2461 tttgataagc ttctggcacc caccagtctt ctcataaacg aatcgtttca ggtgcagcac
     2521 gaggacagga gggagttttt ccagagtcac tcttcgactt atctcaacct cttgtttggt
     2581 ttttgtggta taaccttgga cagattctct tgccaccaag ctctccagtg catcctggac
     2641 tgtgcgtatc ttgtctgact ggatatccaa ctgcaacgtg aaaaatggct gcaaagtggc
     2701 agattctttt gaactctgct ggtaaaccac agacctgatg tgtccaccaa aaatgccggt
     2761 gattggagtc tgaacaaaat ccgcctggcg ggtgacggaa gtcttgttcc gggggcccac
     2821 ttgttcccat tcatcctcgc ttccttcacc ttgttcttcc tgctcttctt cattgaccga
     2881 gtggtttttg gggccgttgg aaatcgtaag tttttcatta cttggtgaga gaagcttctt
     2941 taggttcaac atttcctcat gaagtccatt tagaatgaag cctaagtatt cctcagcatc
     3001 ttcttgtcga cccttttcag acaggcttga cttgttaact gtcaggagtc tgtaaatata
     3061 tgtgggctca aaggcagctc cagggcgaat atccctcacg attttatctc caagagcttg
     3121 tcggggtttt ggaggtactg gcatattagt gaactcattc attagccgaa caaagctgtc
     3181 tatcatgggt gttgacgtac aaggcctttg cactttggaa tacagaggaa tgaacttcat
     3241 caggtggtac atcggcgggc aagcaaccaa tgcctgcagt gtagcattaa tgtagcacca
     3301 gttcccttta ttgatcagcc cacggggttg caacgacact ggtttatgga ttagggttac
     3361 attctccagc aactctgcaa tctttatggc tacaggatcc tctgaaaccg gaacaagccc
     3421 ttctttgact tcaacctgct tttcagaaac caggggagat atggcgggag gggaatactt
     3481 agtttccaca taggccaccg gcgaggagga agagggctta gaatcatgaa agaggctggc
     3541 ccaggacttg ggctggctga caggaagggt gcctgatgca gagcccgtgc cgtcagcagg
     3601 aggtgatgca ctctcgggtt tggttgggtc caagtctatg ctttccgtgg tgtgcaactc
     3661 caccccgttg gtagctgtgc cctcacccga ggattcaagt atttgtccat tagcaactcc
     3721 aaggttttca gtagtatcgg taccaacgca gggctgagcc ccagctgtcc ttgacagggt
     3781 gtctctgcca gcctctgcag ggaagcagga ctgaccaaaa tcagccccgg ggcccccctc
     3841 tggctgccct gcagtcctgg tgtcactgcc gagtgctccg gggaaaggac tgtcaggcac
     3901 aatgtcactg acagagtctg tggagttctg ggggctgtta caagtcctgg gcgtaagtgg
     3961 cggaggcatg tcacccataa attctgcatc ctctgcactg acactgttcg ggactgctga
     4021 attggcatgg ccattgacca gggcttctgt ggagatacta tcatcgccac catctttcaa
     4081 atagctgtaa tatccaggtg gccgcttttt cttcttttta cgctcccttt gtccaagacc
     4141 acctgagaca ccatcatttt ccaaaacttc cgcctccaca ttagaacttc catccaaagc
     4201 gagggcagag cctgggtact ggcagtcgat ggagccatag cttgcttctt tagtgatacc
     4261 atcaggggtt attttggaag ctgtacaacc gagaataaat tcaggggcct gagggttcag
     4321 tgtgcttgaa atactgtagc tgggggttct cggcaaagtg tcactgggtt caatgacttc
     4381 atcgacacca aactcaattc tctgatattc ttgtccatca ggtagtttat ccacagcctg
     4441 tgtgccacac agaactgttc cactgtatgg aggaagctca actgaagatc gaggagtcac
     4501 aaagaattga ttgaattcat cagggctaaa atctccaaaa atatactgcg ggctgtggag
     4561 ggccatgcca acttttttgt acaaacttgt tgatagctta actagccagc ttgggtctcc
     4621 ctatagtgag tcgtattaga tctaattccg gcgcctagag aaggagtgag ggctggataa
     4681 agggaggatc gaggcggggt cgaacgagga ggttcaaggg ggagagacgg ggcggatgga
     4741 ggaagaggag gcggaggctt agggtgtaca aagggcttga cccagggagg ggggtcaaaa
     4801 gccaaggctt cccaggtcac gatgtagggg acctggtctg ggtgtccatg cgggccaggt
     4861 gaaaagacct tgatcttaac ctgggtgatg aggtctcggt taaaggtgcc gtctcgcggc
     4921 catccgacgt taaaggttgg ccattctgca gagcagaagg taacccaacg tctcttcttg
     4981 acatctaccg actggttgtg agcgatccgc tcgacatctt tccagtgacc taaggtcaaa
     5041 cttaagggag tggtaacagt ctggccctaa ttttcagaca aatacagaaa cacagtcaga
     5101 cagagacaac acagaacgat gctgcagcgc tgcagcagac aagacgcgcg gcttcggttc
     5161 caaaccgaaa gcaaaaattc agacggaggc gggaactgtt ttaggttctc gtctcctacc
     5221 agaaccacat atcctgacgg ggtcggattc cacatcgact cccttcctca ggtcgggcca
     5281 caaaaacggc ccccaaagtc cctgggacgt ctcccagggt tgcggccggg tgttcagaac
     5341 tcgtcagttc caccacgggt ccgccagata cagagctagt tagctaacta gtacagacgc
     5401 aggcgcaaac attagatgcc ggcacaaaca cgcacaaaga cagagacaga cacgaaacga
     5461 ccgctggcca gcttacctcc cggcgggggg gtcggtggtc cctgggcagg ggtctccaaa
     5521 tctcggtgga acctccaaat gaaagacccc cgaggtgggc agtcaatcaa tctgaggaga
     5581 ccctcccaag gatcagcgag tccacgattc ggatgcaaac agcaagaggc tttattggga
     5641 atacgggtac ccgggcgacg cagtctatcg gaggactggc gcgccgagtg aggggttgtg
     5701 ggctctttta ttgagctcgg ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat
     5761 tggttagttc aaataaggca cagggtcatt tcaggtcctt ggggcaccct ggaaacatct
     5821 gatggttctc tagaaactgc tgagggcggg accgcatctg gggaccatct gttcttggcc
     5881 ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat tctgctgtct
     5941 ctctgttcct aaccttgatc tgaacttctc tattctcagt tatgtatttt ccatgccttg
     6001 caaaatggcg ttacttaagc tagcttgcca aacctacagg tggggtcttt cattcccccc
     6061 tttttctgga gactaaataa aatcttttat tttatctatg gctcgtactc tataggcttc
     6121 agctggtgat attgttgagt caaaactaga gcctggacca ctgatatcct gtctttaaca
     6181 aattggacta atcgcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
     6241 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
     6301 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
     6361 ttgggcgcca tctccttgca tgcaccattc cttgcgccgt cgttttacaa cgtcgtgact
     6421 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     6481 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     6541 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6601 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     6661 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6721 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6781 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     6841 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6901 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6961 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7021 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     7081 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     7141 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     7201 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     7261 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     7321 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7381 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7441 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7501 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7561 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7621 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7681 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7741 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7801 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7861 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     8161 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaagggga aacgcctggt
     8521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8641 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8701 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8761 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     8821 gttggccgat tcattaatg
//
