LOCUS       sequence-201964-        9287 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 9287)
  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 9287)
  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-USP15
FEATURES             Location/Qualifiers
     source          1..9287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    115..136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(151..156,157..174,175..181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    189..205
                     /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             367..881
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(957..1556)
                     /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(1040..1060)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1537..1556
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1568..2120
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1570..2137
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1730..1749)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1959..1976
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     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(5034..5058)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5074..5100)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5113..5136)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      5136..5145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5158..5574)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5174..5190)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5206..5228)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5641..5982
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             6046..6562
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6762..6781)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6909..6931)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7125..7144)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7244..7266
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7304..7322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7390..7494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7495..7563,7564..8355)
                     /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(7713..7732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8526..9114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9015..9034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9268..9285
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      121 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      181 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      241 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      301 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      361 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      421 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      481 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      541 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      601 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      661 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      721 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
      781 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
      841 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
      901 ctaaataaaa tcttttattt tatcgataag cttggctgca ggtcgacatc gatgcatcag
      961 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     1021 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     1081 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     1141 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     1201 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     1261 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     1321 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     1381 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     1441 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     1501 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt
     1561 gtggccatat tatcatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc
     1621 tagacgtttt tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc
     1681 agatcccata caatggggta ccttctgggc atccttcagc cccttgttga atacgcttga
     1741 ggagagccat ttgactcttt ccacaactat ccaactcaca acgtggcact ggggttgtgc
     1801 cgcctttgca ggtgtatctt atacacgtgg cttttggccg cagaggcacc tgtcgccagg
     1861 tggggggttc cgctgcctgc aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc
     1921 cagaggaact gcttccttca cgacattcaa cagaccttgc attcctttgg cgagagggga
     1981 aagaccccta ggaatgctcg tcaagaagac agggccaggt ttccgggccc tcacattgcc
     2041 aaaagacggc aatatggtgg aaaataacat atagacaaac gcacaccggc cttattccaa
     2101 gcggcttcgg ccagtaacgt tagggggggg gggcggaatt cgttatcgaa ccactttgta
     2161 caagaaagtt gggtattagt tagtgtgcat acagttttca ttttctatat cattgtcatt
     2221 atcattgcta tcttcatcac tttctaatgg gatgccagtg gcagctgaag cacctttagt
     2281 ttctcggtca agaggaaaaa agccagttcc actgaaagtg tcttgtctct ggtagaagag
     2341 tacatatgct gctttggaca caatttggtc ttcagatgca gtggagacac tactgtcatc
     2401 aaaatagtac cattttccat catctttatt ttttgcaaaa gcagtatagt gtcctcctcc
     2461 catccctcca tagtggttgg aaacagcaat cagattatag cggcaaggac ctgcatttgg
     2521 attaattaag aattccgaca tatccaagtc attgatagga aaatcaacta aggtatccaa
     2581 cttgtctctc atgtatcgac tgtaagaaaa tcgcttgaga tgtactacaa gtactggagg
     2641 cagggaccat aaatccaatt tctttgtggc ttgctgatgt tctttacaat tcggacaata
     2701 ccagggatct tcagcaccta gcttttcttt tgttgtaaaa agttcaatgc aatcttttaa
     2761 tttcacaaag ggttttttag gaggtttata ctccacactt tcatgttttt caaagtcctc
     2821 agcagcattt tcatcaaaat atcttttttt caaatcagga tcccaatcca aagcaagaaa
     2881 agatctttca tctagcctaa gctgcctatc atcaaatctt atatgcctgg tatcatcttt
     2941 gatgtagttg atatcagtat tgcctaagtt gttgaactgg aatgtaaaca atcgtttttt
     3001 gtgtcccgtg agttgacctt tgcaagtatc ctcagtacat aatccatttt cagaatcatt
     3061 atctcctcca actgaatctt cagactgact gttttcattc tctgagggaa gttcttgatc
     3121 ctggctggat tcatcatctg gctcatctgt ttccatttca cttggtgagc cttcttcatg
     3181 tatgccattt gggccattcc cattaatatt ttggtcctta cagcagtgta gggatccttc
     3241 agtttcttca gtttcagtag atattttgac atatcggcac attctcaaga gcaggagatt
     3301 ataaagtttg tcttcagtat tgtttcgtgg tacagccata agaaagggct gaccaaaaag
     3361 tgaagaacca gtatggtggg tataactcga gtgtctgaat ttttctctta ggcaaacagg
     3421 aataatcacg tgctctgtat cttctgtcct attgatgtta atttcaaaca cataaatatc
     3481 atcccgttcc ataatactac taaggttttc atccatagcg aatattctgt gaaatctatg
     3541 attgtatata tcagtaacta tcatcttatc tgcaggtatt cctgacaaag cagacaatgc
     3601 tgtacaaaga tctaatatgt ttccaatttt ggggacaacc actttgtact gcataggttt
     3661 ggtaagcgga tccattctaa ctaagtaaac ttccaaggtg cgttcttttt tcatgggcaa
     3721 tggaagtgtc aagtaacaaa aaggatcaaa tgttactgaa atcttagcac actcaggaca
     3781 aactaaagtt gatttgaaaa ggccatgaaa tatatctact atgatagaat catttcgttt
     3841 taaatggttt tcccaggctt cttcggcaac caccttatct ggccttccat ctgcatcttt
     3901 taattgtata tatggttttt tcctaattct attcaaatcc tcatgtaatc catctaatag
     3961 gaaagctaac agttcttgac agtcttgctg ctgatatcca gagaactgag gtgcaaaacg
     4021 tcctacctgt gtcttaaagg ctcttggggt gacgtagcta aactttccag accacatttg
     4081 cttgatcagt tcggcataag atttagctat ttcacctctc attcctaagg gattgtcaaa
     4141 attcagttct tcttgatact tatcattgag gaaatactca gtaagtggag gtgtgttgct
     4201 caaacactga atagctgagt tcatgaaaca cgtatttccc aagttactta ggccacagag
     4261 gcctggctgt tcattgtttc ttccaggttc cgaataatca tagttcttat aagcggtata
     4321 tgatggaaga cagtaatttg agtttttcac attaggagta gaaggacccc ttggccatgt
     4381 tccatcttca tttttctgtt ctatcactaa tacctgtcct tggtataaac cagcatcctg
     4441 aatggtgctg tctggtttat tcagtggttc aaatgtgtta ctcatgtatt tgttccacaa
     4501 tctggtctcc ttttcatctg gaatactgaa gatttttctt atttcctttt caattgtatc
     4561 tattgtgtca gctttgctaa atcttcgagt tacaacatta ttcatgtttc cattttcaca
     4621 tagcttcaat tctgtgagat atacttctac tttgcagtgc tttacaaaca taccctgttc
     4681 aaccaccttt cgtgctattg gctcttgacc ttccatcaat gtgtaccagc tgacaagttt
     4741 attccaacct tcagttggca acagtatgta atccaattca tcaataagat gttccttaag
     4801 tgactgggca tcaccatctt tgagaagtcc agagttatca atgggtccag gatacacatt
     4861 ttgatctccc atctggtatt tgtcccaact gtcaaagcca acatattttt tccactgttt
     4921 gaaccagcga ctatcgacta ggtaccaggt gtcccctttc cggagcgagg ttttgagcag
     4981 cgtcgcgatg tcagaccgct gggtgtccag atccgccgct ccgccttccg ccatgccaac
     5041 ttttttgtac aaacttgttt gataacctcg agcagcgtaa tctggaacat cgtatgggta
     5101 tcctccatcg agcttgtcat cgtcatcctt gtagtccatg gtggcagatc taattccggc
     5161 gcctagagaa ggagtgaggg ctggataaag ggaggatcga ggcggggtcg aacgaggagg
     5221 ttcaaggggg agagacgggg cggatggagg aagaggaggc ggaggcttag ggtgtacaaa
     5281 gggcttgacc cagggagggg ggtcaaaagc caaggcttcc caggtcacga tgtaggggac
     5341 ctggtctggg tgtccatgcg ggccaggtga aaagaccttg atcttaacct gggtgatgag
     5401 gtctcggtta aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc attctgcaga
     5461 gcagaaggta acccaacgtc tcttcttgac atctaccgac tggttgtgag cgatccgctc
     5521 gacatctttc cagtgaccta aggtcaaact taagggagtg gtaacagtct ggccctaatt
     5581 ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc tgcagcgctg
     5641 cagcagacaa gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag acggaggcgg
     5701 gaactgtttt aggttctcgt ctcctaccag aaccacatat cctgacgggg tcggattcca
     5761 catcgactcc cttcctcagg tcgggccaca aaaacggccc ccaaagtccc tgggacgtct
     5821 cccagggttg cggccgggtg ttcagaactc gtcagttcca ccacgggtcc gccagataca
     5881 gagctagtta gctaactagt acagacgcag gcgcaaacat tagatgccgg cacaaacacg
     5941 cacaaagaca gagacagaca cgaaacgacc gctggccagc ttacctcccg gcgggggggt
     6001 cggtggtccc tgggcagggg tctccaaatc tcggtggaac ctccaaatga aagacccccg
     6061 aggtgggcag tcaatcaatc tgaggagacc ctcccaagga tcagcgagtc cacgattcgg
     6121 atgcaaacag caagaggctt tattgggaat acgggtaccc gggcgacgca gtctatcgga
     6181 ggactggcgc gccgagtgag gggttgtggg ctcttttatt gagctcgggg agcagaagcg
     6241 cgcgaacaga agcgagaagc gaactgattg gttagttcaa ataaggcaca gggtcatttc
     6301 aggtccttgg ggcaccctgg aaacatctga tggttctcta gaaactgctg agggcgggac
     6361 cgcatctggg gaccatctgt tcttggccct gagccggggc aggaactgct taccacagat
     6421 atcctgtttg gcccatattc tgctgtctct ctgttcctaa ccttgatctg aacttctcta
     6481 ttctcagtta tgtattttcc atgccttgca aaatggcgtt acttaagcta gcttgccaaa
     6541 cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa tcttttattt
     6601 tatctatggc tcgtactcta taggcttcag ctggtgatat tgttgagtca aaactagagc
     6661 ctggaccact gatatcctgt ctttaacaaa ttggactaat cgcctctacg ccggacgcat
     6721 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     6781 cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat
     6841 ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct
     6901 tgcgccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     6961 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     7021 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     7081 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     7141 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     7201 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     7261 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     7321 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     7381 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     7621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     7681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     7741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     7801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     7861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     8041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     8221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     8341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     8401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     8461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     8521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     8581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     8641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     8701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     8761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     8821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     8881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     9001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     9181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     9241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgagga
//
