LOCUS       sequence-313904-        7988 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 7988)
  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 7988)
  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-USP3
FEATURES             Location/Qualifiers
     source          1..7988
                     /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(2102..2126)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2139..2706
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2156..2708
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2300..2317)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2527..2546
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2720..3319
                     /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(2720..2739)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3216..3236
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3395..3909
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4071..4087
                     /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(4095..4101,4102..4119,4120..4125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4140..4161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4278..4295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4449..5037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4529..4548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5208..5999,6000..6068))
                     /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     5831..5850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6069..6173)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6241..6259
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6297..6319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6419..6438
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6632..6654
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6782..6801
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7001..7517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7581..7922
                     /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 atggagtgtc cacacctgag ctccagcgtc tgcattgctc cggactcagc caagttcccc
      601 aacggctccc cgtcgtcctg gtgctgcagc gtgtgccggt ccaacaaaag cccttgggtc
      661 tgtttgactt gttcaagtgt ccactgtgga aggtatgtga atggccatgc aaaaaaacat
      721 tatgaagatg cacaagtacc tttaaccaac cataagaaat cagaaaagca agataaagtt
      781 cagcacacag tatgtatgga ttgcagtagc tacagtacat actgttatcg ctgtgatgat
      841 tttgtggtta atgacaccaa gctgggactg gtacagaaag tcagagaaca cttacagaac
      901 ttggaaaact cagctttcac agctgacagg cataagaaaa gaaaactttt ggaaaactca
      961 acactaaaca gcaagttatt aaaagtaaat ggaagcacca ctgccatttg tgccacaggc
     1021 cttcggaatt tggggaacac atgtttcatg aatgccatcc ttcagtcact cagtaacatt
     1081 gagcagtttt gctgttattt caaagaactg cccgccgtgg agttaaggaa tgggaaaaca
     1141 gcaggaaggc ggacatacca caccaggagc caaggggata acaatgtgtc tttggtagaa
     1201 gagtttagaa agacactctg tgctttatgg caaggcagcc agactgcatt tagcccagag
     1261 tccttatttt atgttgtttg gaagattatg ccaaacttta ggggctatca acagcaggac
     1321 gcccatgaat tcatgcgcta ccttttggac cacctacact tggaacttca gggcggtttc
     1381 aacggtgttt cccgctcagc aattctgcag gagaattcta ctctgtctgc aagtaacaag
     1441 tgttgcataa atggagcatc tactgttgtc acggctatat tcggaggcat tctccaaaat
     1501 gaggttaact gcctcatatg tgggacagaa tctagaaagt ttgatccatt cctagacctt
     1561 tcattagata ttccaagtca gttcagaagt aagcgctcta agaatcaaga aaatggacca
     1621 gtttgttcgt tacgagattg tcttcgcagt tttaccgact tagaagaact tgatgagaca
     1681 gagttatata tgtgccataa atgcaaaaag aaacaaaagt ccacaaaaaa gttttggatt
     1741 caaaaactac ccaaggtgct atgcttacat ttgaaaagat ttcattggac agcatattta
     1801 agaaataaag ttgatacata cgtagaattt ccactgagag gcctagacat gaaatgctac
     1861 ttactagagc ctgagaacag tggcccggag agctgcctgt atgacctcgc cgctgtggtg
     1921 gtgcaccatg gttccggggt tggttctgga cattacacag catacgcaac tcacgaaggc
     1981 cgctggttcc acttcaatga cagtactgta acactgactg acgaggagac tgtggtgaag
     2041 gcgaaggcct acatcctttt ctacgtggaa caccaggcca aagctggatc ggataaactt
     2101 taaccaactt tcttgtacaa agtggttcga taacgaattc cgcccccccc ccctaacgtt
     2161 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     2221 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2281 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2341 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2401 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2461 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2521 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2581 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     2641 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     2701 tgataatatg gccacaacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     2761 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     2821 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     2881 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     2941 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3001 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3061 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     3121 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     3181 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     3241 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     3301 ccgcaagccc ggtgcctgat gcatcgatgt cgacctgcag ccaagcttat cgataaaata
     3361 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     3421 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     3481 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     3541 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     3601 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3661 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3721 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3781 tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     3841 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     3901 gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga
     3961 atattaagta atcctgactc aattagccac tgttttgaat ccacatactc caatactcct
     4021 gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg
     4081 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4141 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4201 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4261 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4321 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4381 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4441 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4501 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4561 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4621 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4681 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4741 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4801 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4861 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4921 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4981 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5041 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5101 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5161 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5221 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5281 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5341 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5401 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5461 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5521 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5581 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5641 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5701 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5761 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5821 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5881 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5941 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6001 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6061 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6121 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6181 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6241 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     6301 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     6361 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     6421 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     6481 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     6541 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6601 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
     6661 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6721 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6781 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6841 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
     6901 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     6961 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     7021 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     7081 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     7141 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     7201 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     7261 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     7321 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     7381 cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc
     7441 atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac
     7501 ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac
     7561 cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg
     7621 tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc
     7681 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
     7741 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
     7801 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
     7861 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
     7921 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
     7981 aaaattag
//
