LOCUS       sequence-181044-        8615 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 8615)
  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 8615)
  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-BAP1
FEATURES             Location/Qualifiers
     source          1..8615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     330..349
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             549..1065
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1129..1470
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1537..1953
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1883..1905
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1921..1937
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1966..1975
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1975..1998
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2011..2037
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2053..2077
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    3265..3276
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(4265..4289)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4302..4869
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4319..4871
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4463..4480)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4690..4709
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4883..5482
                     /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(4883..4902)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5379..5399
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5558..6072
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6234..6250
                     /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(6258..6264,6265..6282,6283..6288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6303..6324
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6441..6458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6612..7200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6692..6711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7371..8162,8163..8231))
                     /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     7994..8013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8232..8336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8404..8422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8460..8482)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8582..8601
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
       61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      181 ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga gatggcgccc
      241 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
      301 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
      361 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg attagtccaa
      421 tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca atatcaccag
      481 ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag tctccagaaa
      541 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt
      601 tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag
      661 agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
      721 agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca
      781 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
      841 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
      901 cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt
      961 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
     1021 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
     1081 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
     1141 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
     1201 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     1261 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
     1321 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
     1381 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     1441 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
     1501 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
     1561 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
     1621 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
     1681 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
     1741 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
     1801 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
     1861 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
     1921 ctttatccag ccctcactcc ttctctaggc gccggaatta gatctgccac catggactac
     1981 aaggatgacg atgacaagct cgatggagga tacccatacg atgttccaga ttacgctgct
     2041 cgaggttatc aaacaagttt gtacaaaaaa gttggcatga ataagggctg gctggagctg
     2101 gagagcgacc caggcctctt caccctgctc gtggaagatt tcggtgtcaa gggggtgcaa
     2161 gtggaggaga tctacgacct tcagagcaaa tgtcagggcc ctgtatatgg atttatcttc
     2221 ctgttcaaat ggatcgaaga gcgccggtcc cggcgaaagg tctctacctt ggtggatgat
     2281 acgtccgtga ttgatgatga tattgtgaat aacatgttct ttgcccacca gctgataccc
     2341 aactcttgtg caactcatgc cttgctgagc gtgctcctga actgcagcag cgtggacctg
     2401 ggacccaccc tgagtcgcat gaaggacttc accaagggtt ttagccctga gagcaaagga
     2461 tatgcgattg gcaatgcccc ggagttggcc aaggcccata atagccatgc caggcccgag
     2521 ccacgccacc tccctgagaa gcagaatggc cttagtgcag tgcggaccat ggaggcattc
     2581 cactttgtca gctatgtgcc tatcacaggc cggctctttg agctggatgg gctgaaggtc
     2641 taccccattg accatgggcc ctggggggag gacgaggagt ggacagacaa ggcccggcgg
     2701 gtcatcatgg agcgtatcgg cctcgccact gcaggggagc cctaccacga catccgcttc
     2761 aacctgatgg cagtggtgcc cgaccgcagg atcaagtatg aggccaggct gcatgtgctg
     2821 aaggtgaacc gtcagacagt actagaggct ctgcagcagc tgataagagt aacacagcca
     2881 gagctgattc agacccacaa gtctcaagag tcacagctgc ctgaggagtc caagtcagcc
     2941 agcaacaagt ccccgctggt gctggaagca aacagggccc ctgcagcctc tgagggcaac
     3001 cacacagatg gtgcagagga ggcggctggt tcatgcgcac aagccccatc ccacagccct
     3061 cccaacaaac ccaagctagt ggtgaagcct ccaggcagca gcctcaatgg ggttcacccc
     3121 aaccccactc ccattgtcca gcggctgccg gcctttctag acaatcacaa ttatgccaag
     3181 tcccccatgc aggaggaaga agacctggcg gcaggtgtgg gccgcagccg agttccagtc
     3241 cgcccacccc agcagtactc agatgatgag gatgactatg aggatgacga ggaggatgac
     3301 gtgcagaaca ccaactctgc ccttaggtat aaggggaagg gaacagggaa gccaggggca
     3361 ttgagcggtt ctgctgatgg gcaactgtca gtgctgcagc ccaacaccat caacgtcttg
     3421 gctgagaagc tcaaagagtc ccagaaggac ctctcaattc ctctgtccat caagactagc
     3481 agcggggctg ggagtccggc tgtggcagtg cccacacact cgcagccctc acccaccccc
     3541 agcaatgaga gtacagacac ggcctctgag atcggcagtg ctttcaactc gccactgcgc
     3601 tcgcctatcc gctcagccaa cccgacgcgg ccctccagcc ctgtcacctc ccacatctcc
     3661 aaggtgcttt ttggagagga tgacagcctg ctgcgtgttg actgcatacg ctacaaccgt
     3721 gctgtccgtg atctgggtcc tgtcatcagc acaggcctgc tgcacctggc tgaggatggg
     3781 gtgctgagtc ccctggcgct gacagagggt gggaagggtt cctcgccctc catcagacca
     3841 atccaaggca gccaggggtc cagcagccca gtggagaagg aggtcgtgga agccacggac
     3901 agcagagaga agacggggat ggtgaggcct ggcgagccct tgagtgggga gaaatactca
     3961 cccaaggagc tgctggcact gctgaagtgt gtggaggctg agattgcaaa ctatgaggcg
     4021 tgcctcaagg aggaggtaga gaagaggaag aagttcaaga ttgatgacca gagaaggacc
     4081 cacaactacg atgagttcat ctgcaccttt atctccatgc tggctcagga aggcatgctg
     4141 gccaacctag tggagcagaa catctccgtg cggcggcgcc aaggggtcag catcggccgg
     4201 ctccacaagc agcggaagcc tgaccggcgg aaacgctctc gcccctacaa ggccaagcgc
     4261 cagtgcccaa ctttcttgta caaagtggtt cgataacgaa ttccgccccc cccccctaac
     4321 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     4381 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     4441 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     4501 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     4561 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     4621 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     4681 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     4741 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4801 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     4861 cgatgataat atggccacaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4921 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4981 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     5041 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5101 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5161 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5221 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5281 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5341 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5401 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5461 gacccgcaag cccggtgcct gatgcatcga tgtcgacctg cagccaagct tatcgataaa
     5521 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     5581 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     5641 gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat
     5701 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     5761 cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     5821 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     5881 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc
     5941 cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     6001 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     6061 ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt
     6121 cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact
     6181 cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat
     6241 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6301 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6361 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6781 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7021 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7681 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7981 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8341 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     8401 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     8461 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     8521 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     8581 gcggcatcag agcagattgt actgagagtg cacca
//
