LOCUS       sequence-327661-        9032 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 9032)
  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 9032)
  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-USP13
FEATURES             Location/Qualifiers
     source          1..9032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             65..579
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    741..757
                     /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(765..771,772..789,790..795))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    810..831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(948..965)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1119..1707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1199..1218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1878..2669,2670..2738))
                     /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     2501..2520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2739..2843)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2911..2929
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2967..2989)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3089..3108
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3302..3324
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3452..3471
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             3671..4187
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    4251..4592
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             4659..5075
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5005..5027
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5043..5059
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      5088..5097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5097..5120
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5133..5159
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    5175..5199
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7804..7828)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    7841..8408
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7858..8410
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8002..8019)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8229..8248
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8422..9021
                     /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(8422..8441)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8918..8938
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgacctgcag ccaagcttat cgataaaata aaagatttta tttagtctcc agaaaaaggg
       61 gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa
      121 ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg aacagagaga
      181 cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
      241 caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga
      301 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
      361 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
      421 aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc
      481 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
      541 ctctgagtga ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt
      601 ggagaacaac attctgaggg taggagtcga atattaagta atcctgactc aattagccac
      661 tgttttgaat ccacatactc caatactcct gaaatagttc attatggaca gcgcagaaag
      721 agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      781 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      841 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      901 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
      961 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1021 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1081 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1141 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1201 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1261 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1321 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1381 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1441 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1501 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1561 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1621 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1681 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1741 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1801 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1861 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1921 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1981 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2041 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2101 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2161 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2221 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2281 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2341 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2401 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2461 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2521 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2581 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2641 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2701 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2761 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2821 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     2881 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     2941 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3001 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3061 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3121 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     3181 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3241 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3301 tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc
     3361 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     3421 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     3481 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc
     3541 aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc
     3601 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga
     3661 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     3721 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     3781 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     3841 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     3901 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     3961 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     4021 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     4081 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg
     4141 agggtctcct cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc
     4201 gagatttgga gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca
     4261 gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc
     4321 gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
     4381 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
     4441 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
     4501 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     4561 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
     4621 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     4681 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     4741 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     4801 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     4861 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     4921 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     4981 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     5041 ccctttatcc agccctcact ccttctctag gcgccggaat tagatctgcc accatggact
     5101 acaaggatga cgatgacaag ctcgatggag gatacccata cgatgttcca gattacgctg
     5161 ctcgaggtta tcaaacaagt ttgtacaaaa aagttggcgc gcccaccatg cagcgccggg
     5221 gcgccctgtt cggcatgccg ggcggcagcg gaggcaggaa gatggctgca ggagacatcg
     5281 gcgagctgct agtgccccac atgcccacga tccgcgtgcc caggtccggc gacagggtct
     5341 acaagaacga gtgcgccttc tcctacgact ctcccaattc tgaaggtgga ctctatgtat
     5401 gcatgaatac atttttggcc tttggaaggg aacatgttga aagacatttt cgaaaaactg
     5461 gacagagtgt atacatgcac ctgaaaagac atgtgcgaga gaaggtaaga ggggcgtctg
     5521 gtggagcgtt accaaaaagg aggaattcca agattttttt agatctagat actgatgacg
     5581 atttaaatag cgacgattat gaatatgaag atgaagccaa acttgttata ttcccagatc
     5641 actatgaaat agcactacca aatattgagg agttaccagc cctggtaaca attgcttgtg
     5701 atgcagttct cagctcaaaa tctccataca gaaagcagga cccagacacg tgggaaaatg
     5761 aattgccagt atctaaatat gccaacaacc tcacccagct ggacaatgga gtcaggattc
     5821 ctccaagtgg ttggaagtgt gccagatgcg acctgcgaga aaacctctgg ttgaatctga
     5881 ctgacggctc tgtcctgtgt ggaaagtggt tctttgacag ctctgggggc aacgggcatg
     5941 cgctggagca ttacagagac atgggctacc cactagccgt gaaactggga accatcactc
     6001 ctgacggggc agatgtttat tcttttcaag aagaagaacc tgttttggat cctcatttgg
     6061 ccaagcactt agcgcatttt ggaattgata tgcttcatat gcatgggaca gagaatgggc
     6121 tccaggacaa tgacatcaag ctgagggtca gtgagtggga agtgatccag gagtcgggca
     6181 cgaaactgaa gccaatgtat ggtcctggct acacgggtct gaagaacctg ggcaacagct
     6241 gctatctcag ctctgtcatg caggccatct tcagcatccc agaattccag agagcgtatg
     6301 taggaaacct tcccagaata tttgactact cgcctttaga tccaacacaa gatttcaaca
     6361 cacagatgac taagttagga catggccttc tctcaggcca gtattcaaag cctccggtga
     6421 aatctgaact cattgaacag gtgatgaagg aggagcacaa gccacagcag aacgggatct
     6481 ctccgcgcat gtttaaggcc tttgtaagca agagccaccc ggaattctcc tctaacaggc
     6541 agcaagatgc ccaggaattc ttcttgcacc tggtgaatct agtagagagg aaccgcatcg
     6601 gctcagaaaa cccaagcgat gtttttcgtt ttttggtgga agaacgcatt cagtgctgtc
     6661 agacccggaa agtccgctac acggagaggg tggattacct gatgcagtta cctgtggcca
     6721 tggaggcggc aaccaacaag gatgaactga tcgcttatga actaacgaga agggaagcag
     6781 aagcaaacag aagacccctt cctgagttgg tacgtgccaa gataccattt agtgcctgcc
     6841 ttcaggcctt ctctgaacca gaaaatgttg atgatttctg gagcagtgcc ctacaagcaa
     6901 agtctgcggg tgtgaaaaca tctcgctttg cttcattccc tgaatacttg gtagtgcaga
     6961 taaagaagtt cacttttggt cttgactggg ttcccaaaaa atttgatgtt tctattgata
     7021 tgccagacct acttgatatc aaccatctcc gagccagggg gttacagcca ggagaggaag
     7081 aacttccaga catcagcccc cccatagtca ttcctgatga ctcaaaagat cgcctgatga
     7141 accaattgat agacccatca gacatcgatg agtcatcagt gatgcagctg gccgagatgg
     7201 gtttcccgct ggaagcatgt cgcaaggctg tgtacttcac tggaaatatg ggcgccgagg
     7261 tggccttcaa ctggatcatt gttcacatgg aagagccaga ttttgctgag ccgctgacca
     7321 tgcctggtta tggaggggca gcttctgctg gagcctctgt ttttggtgct tctggactgg
     7381 ataaccaacc tccagaggaa atcgtagcta tcatcacctc catgggattt cagcgaaatc
     7441 aggctattca ggcactacga gcaacgaata ataacctgga aagagcactg gattggatct
     7501 ttagccaccc tgagtttgaa gaagacagtg attttgtgat tgagatggag aataatgcca
     7561 atgcaaacat tatttctgag gccaagcccg aaggacctag agtcaaggat ggatctggaa
     7621 catatgagct atttgcattc atcagtcaca tgggaacatc cacaatgagt ggtcattaca
     7681 tttgccatat caaaaaggaa ggaagatggg tgatttacaa tgaccacaaa gtttgtgcct
     7741 cagaaaggcc ccctaaagac ctgggctaca tgtactttta ccgcaggata ccaagcttaa
     7801 ttaatccaac tttcttgtac aaagtggttc gataacgaat tccgcccccc ccccctaacg
     7861 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     7921 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     7981 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     8041 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     8101 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     8161 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     8221 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     8281 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     8341 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     8401 gatgataata tggccacaac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     8461 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     8521 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     8581 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     8641 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     8701 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     8761 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     8821 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     8881 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     8941 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     9001 acccgcaagc ccggtgcctg atgcatcgat gt
//
