LOCUS       sequence-132791-        7376 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 7376)
  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 7376)
  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-BRCC3
FEATURES             Location/Qualifiers
     source          1..7376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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             359..873
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(949..1548)
                     /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(1032..1052)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1529..1548
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1560..2112
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1562..2129
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1722..1741)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1951..1968
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    2142..2166
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3115..3139)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3155..3181)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3194..3217)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3217..3226
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3239..3655)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3255..3271)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3287..3309)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3722..4063
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4127..4643
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4843..4862)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4990..5012)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5206..5225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5325..5347
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5385..5403)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5471..5575
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5576..5644,5645..6436)
                     /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(5794..5813)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6607..7195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7096..7115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7349..7366
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
      241 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
      301 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
      361 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      421 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
      481 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
      541 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      601 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      661 tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      721 cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc
      781 tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc
      841 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
      901 aatcttttat tttatcgata agcttggctg caggtcgaca tcgatgcatc aggcaccggg
      961 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     1021 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     1081 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     1141 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     1201 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     1261 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     1321 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     1381 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac
     1441 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     1501 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg ttgtggccat
     1561 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt
     1621 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca
     1681 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc
     1741 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg
     1801 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt
     1861 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa
     1921 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc
     1981 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg
     2041 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc
     2101 ggccagtaac gttagggggg gggggcggaa ttcgttatcg aaccactttg tacaagaaag
     2161 ttgggtattc tagagaagaa agttcttgca taagctcttc cttttcttgt tgtaattcct
     2221 gcaaatgctg ttggttttgc tccagtctgt cctccaacca ctgtaggaga ggcccgctga
     2281 ctgccgacat ctgactgcac agattcttgg taaacactga gccattatgg atcttggtta
     2341 ctgagtccag atgtgtaagg ctgtggatcc tcctatacgc atcctgctcc tcctggcaca
     2401 ggatcttggg cagctctact gctgattcaa ggcacacttt cccgatagtg acatgaggta
     2461 caatatggat tgggatttcg attctctcat acctttcctc ttccttctgg ccctcaatgg
     2521 agatgtgctg gctggagctc cagaagtctc gtggaccatg aagggactct gaactctttt
     2581 gggcctgtat ggattggaag caagtgtaga gtacccggcc agtctttgtg ttcttatctt
     2641 ctatgaaaca ggaaaaaata agtcctacaa agccttgatc catcatctgg tacatggctt
     2701 gtgtgcgaac atcaacatgt gaaggccaaa cagttatatg aggatgggaa tgataccagc
     2761 ccacaactct catggggcgg cctgtcagtt cagccaacct ctctgcctct gttgaagctg
     2821 cagacagctg ctctggagaa atttctactc ggtccttcct cttatcagaa cgtcgtaaga
     2881 tgatgacaga atgaatgtga acaattctga cggcatcaac cttttcagca actgtgcgca
     2941 tttcagttcc agtatatgca aatttggagt cactccttgt atcatcgttc aactccccta
     3001 tgcacagccc cattacttcc tccttctctg tgctcagagc gtggttgaga caaacgagga
     3061 aagcgtcaga ctcgagatga accgcctgca ccgcctgcac cacctgcacc gccatgccaa
     3121 cttttttgta caaacttgtt tgataacctc gagcagcgta atctggaaca tcgtatgggt
     3181 atcctccatc gagcttgtca tcgtcatcct tgtagtccat ggtggcagat ctaattccgg
     3241 cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc gaacgaggag
     3301 gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta gggtgtacaa
     3361 agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga
     3421 cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc tgggtgatga
     3481 ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc cattctgcag
     3541 agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga gcgatccgct
     3601 cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc tggccctaat
     3661 tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg ctgcagcgct
     3721 gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag caaaaattca gacggaggcg
     3781 ggaactgttt taggttctcg tctcctacca gaaccacata tcctgacggg gtcggattcc
     3841 acatcgactc ccttcctcag gtcgggccac aaaaacggcc cccaaagtcc ctgggacgtc
     3901 tcccagggtt gcggccgggt gttcagaact cgtcagttcc accacgggtc cgccagatac
     3961 agagctagtt agctaactag tacagacgca ggcgcaaaca ttagatgccg gcacaaacac
     4021 gcacaaagac agagacagac acgaaacgac cgctggccag cttacctccc ggcggggggg
     4081 tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa cctccaaatg aaagaccccc
     4141 gaggtgggca gtcaatcaat ctgaggagac cctcccaagg atcagcgagt ccacgattcg
     4201 gatgcaaaca gcaagaggct ttattgggaa tacgggtacc cgggcgacgc agtctatcgg
     4261 aggactggcg cgccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc
     4321 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt
     4381 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggcggga
     4441 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
     4501 tatcctgttt ggcccatatt ctgctgtctc tctgttccta accttgatct gaacttctct
     4561 attctcagtt atgtattttc catgccttgc aaaatggcgt tacttaagct agcttgccaa
     4621 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt
     4681 ttatctatgg ctcgtactct ataggcttca gctggtgata ttgttgagtc aaaactagag
     4741 cctggaccac tgatatcctg tctttaacaa attggactaa tcgcctctac gccggacgca
     4801 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     4861 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     4921 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     4981 ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5041 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5101 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     5161 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     5221 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5281 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5341 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5401 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5461 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5521 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5581 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5641 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5701 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5761 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5821 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5881 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5941 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6001 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6061 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6121 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6181 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6241 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6301 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6361 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6421 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6481 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6541 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6601 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6661 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6721 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6781 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6841 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6901 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6961 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7021 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7081 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7141 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7201 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7261 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7321 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag
//
