LOCUS       sequence-132788-        7541 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 7541)
  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 7541)
  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-ATXN3
FEATURES             Location/Qualifiers
     source          1..7541
                     /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(3280..3304)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3320..3346)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3359..3382)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3382..3391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3404..3820)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3420..3436)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3452..3474)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3887..4228
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4292..4808
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5008..5027)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5155..5177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5371..5390)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5490..5512
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5550..5568)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5636..5740
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5741..5809,5810..6601)
                     /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(5959..5978)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6772..7360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7261..7280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7514..7531
                     /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 ttgggtatta tttttttcct tctgttttca aatcatttct gacagtttct aaagacatgg
     2221 tcacagctgc ctgaagcatg tcttcttcac tcatagcatc acctagatca ctcccaagtg
     2281 ctcctgaact ggtggctggc ctttcacatg gatgtgaact ctgtcctgat aggtccccct
     2341 gctgctgctg ctgctgctgc tgctgctgct gctgctgctg ctgctgctgt tgctgctttt
     2401 gctgctgttt ttcaaagtag gcttctcgtc tcttccgaag ctcttctgaa gtaagatttg
     2461 tacctgatgt ctgtgtcata tcttgagata tgtttctgga actaccttgc atacttagct
     2521 gaatagccct gcggagatct gcttcctcat cttccatgtc aatttcttgg cgacttagtg
     2581 ccagagccct ctgcaaatcc tcctcatctt cgtctaacat tcctgagcca tcatttgctt
     2641 ctaacactcg ttccaggtct gttttatgga ctctttgctc ttttagttgt gctaattctt
     2701 ctccaataag ttttggtcga tgcatctgtt ggaccctaat catctgcagg agttggtcag
     2761 cttcgcaatc tggcagatca cccttaacga caaatataga ataaccttcc tgttgtaatt
     2821 gagccaagaa aagtgcaaga tatgtatctg atattaattc tggacccgtc aagagagaat
     2881 tcaagttaaa ccactgtttt cctaattttc taactgtaaa ccagtgttcc ttataattgc
     2941 atataaatga tctttcattt ataggatcga tcctgagcct ctgatactct ggactgttga
     3001 acaggattag ttctaaaccc caaactttca aggcattgct tataacctga atagagaaaa
     3061 aaccactgtc atccatattt ccagaaggct gctgtaaaaa cgtgcgataa tcttcactag
     3121 taactcctcc ttctgccatt ctcatcctct cctcctcatc cagctgatgt gcaattgagg
     3181 ataattccac ggggctaaaa tattctcctt gcaataagtt attcaggcaa tgttgagcac
     3241 aaagtgagcc ttcttgtttc tcgtggaaga tggactccat gccaactttt ttgtacaaac
     3301 ttgtttgata acctcgagca gcgtaatctg gaacatcgta tgggtatcct ccatcgagct
     3361 tgtcatcgtc atccttgtag tccatggtgg cagatctaat tccggcgcct agagaaggag
     3421 tgagggctgg ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag
     3481 acggggcgga tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg
     3541 gaggggggtc aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc
     3601 catgcgggcc aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg
     3661 tgccgtctcg cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc
     3721 aacgtctctt cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt
     3781 gacctaaggt caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca
     3841 gaaacacagt cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg
     3901 cgcggcttcg gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt
     3961 tctcgtctcc taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc
     4021 ctcaggtcgg gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc
     4081 cgggtgttca gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta
     4141 actagtacag acgcaggcgc aaacattaga tgccggcaca aacacgcaca aagacagaga
     4201 cagacacgaa acgaccgctg gccagcttac ctcccggcgg gggggtcggt ggtccctggg
     4261 caggggtctc caaatctcgg tggaacctcc aaatgaaaga cccccgaggt gggcagtcaa
     4321 tcaatctgag gagaccctcc caaggatcag cgagtccacg attcggatgc aaacagcaag
     4381 aggctttatt gggaatacgg gtacccgggc gacgcagtct atcggaggac tggcgcgccg
     4441 agtgaggggt tgtgggctct tttattgagc tcggggagca gaagcgcgcg aacagaagcg
     4501 agaagcgaac tgattggtta gttcaaataa ggcacagggt catttcaggt ccttggggca
     4561 ccctggaaac atctgatggt tctctagaaa ctgctgaggg cgggaccgca tctggggacc
     4621 atctgttctt ggccctgagc cggggcagga actgcttacc acagatatcc tgtttggccc
     4681 atattctgct gtctctctgt tcctaacctt gatctgaact tctctattct cagttatgta
     4741 ttttccatgc cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt
     4801 ctttcattcc cccctttttc tggagactaa ataaaatctt ttattttatc tatggctcgt
     4861 actctatagg cttcagctgg tgatattgtt gagtcaaaac tagagcctgg accactgata
     4921 tcctgtcttt aacaaattgg actaatcgcc tctacgccgg acgcatcgtg gccggcatca
     4981 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     5041 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     5101 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg ccgtcgtttt
     5161 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5221 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5281 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5341 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5401 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5461 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5521 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5581 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5641 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5701 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5761 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5821 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5881 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5941 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6001 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6061 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6121 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6181 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6241 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6301 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6361 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6421 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6481 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6541 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6601 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6661 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6721 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6781 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6841 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6901 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6961 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7021 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7081 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7141 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7201 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7261 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7321 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7381 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7441 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7501 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
