LOCUS       sequence-313766-        7133 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 7133)
  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 7133)
  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-OTUB2
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      430..439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             439..462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             475..501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    517..541
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1247..1271)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1284..1851
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1301..1853
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1445..1462)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1672..1691
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1865..2464
                     /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(1865..1884)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2361..2381
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2540..3054
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3216..3232
                     /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(3240..3246,3247..3264,3265..3270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3285..3306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3423..3440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3594..4182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3674..3693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4353..5144,5145..5213))
                     /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     4976..4995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5214..5318)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5386..5404
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5442..5464)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5564..5583
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5777..5799
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5927..5946
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6146..6662
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6726..7067
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
      421 attagatctg ccaccatgga ctacaaggat gacgatgaca agctcgatgg aggataccca
      481 tacgatgttc cagattacgc tgctcgaggt tatcaaacaa gtttgtacaa aaaagttggc
      541 atgagtgaaa catctttcaa cctaatatca gaaaaatgtg acattctatc cattcttcgg
      601 gaccatcctg aaaacaggat ttaccggagg aaaatcgagg aactcagcaa aaggttcacc
      661 gccatccgca agaccaaagg ggatgggaac tgcttctaca gggccttggg ctattcctac
      721 ctggagtccc tgctggggaa gagcagggag atcttcaagt tcaaagaacg cgtactgcag
      781 accccaaatg accttctggc tgctggcttt gaggagcaca agttcagaaa cttcttcaat
      841 gctttttaca gtgtggtgga actggtagag aaggacggct cagtgtccag cctgctgaag
      901 gtgttcaacg accagagtgc ctcggaccac atcgtgcagt tcctgcgcct gctcacgtcg
      961 gccttcatca ggaaccgagc agacttcttc cggcacttca ttgatgagga gatggacatc
     1021 aaagacttct gcactcacga agtagagccc atggccacgg agtgtgacca catccagatc
     1081 acggcgttgt cgcaggccct gagcattgcc ctgcaagtgg agtacgtgga cgagatggat
     1141 accgccctga accaccacgt gttccctgag gccgccaccc cttccgttta cctgctctat
     1201 aaaacatccc actacaacat cctttatgca gccgataaac attgataccc aactttcttg
     1261 tacaaagtgg ttcgataacg aattccgccc ccccccccta acgttactgg ccgaagccgc
     1321 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     1381 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     1441 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     1501 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     1561 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     1621 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     1681 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     1741 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     1801 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     1861 aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     1921 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     1981 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     2041 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     2101 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     2161 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     2221 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     2281 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     2341 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     2401 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     2461 ctgatgcatc gatgtcgacc tgcagccaag cttatcgata aaataaaaga ttttatttag
     2521 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     2581 aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg
     2641 ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct
     2701 gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc
     2761 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     2821 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     2881 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc
     2941 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     3001 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa
     3061 cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct
     3121 gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat
     3181 ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac
     3241 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3301 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3361 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3421 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3481 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3541 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3601 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3661 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3721 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3781 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3841 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3901 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3961 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4021 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4081 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4141 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4201 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4261 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4321 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4381 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4441 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4501 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4561 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4621 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4681 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4741 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4801 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4861 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4921 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4981 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5041 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5101 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5161 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5221 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5281 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5341 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5401 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     5461 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     5521 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     5581 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5641 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5701 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5761 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat
     5821 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     5881 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     5941 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6001 agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     6061 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     6121 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     6181 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     6241 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     6301 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     6361 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     6421 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     6481 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     6541 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac
     6601 tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca
     6661 tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg
     6721 gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg
     6781 gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga
     6841 cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg
     6901 actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc
     6961 cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     7021 aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca
     7081 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tag
//
