LOCUS       sequence-128527-        8653 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 8653)
  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 8653)
  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-OTUD5
FEATURES             Location/Qualifiers
     source          1..8653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1420..1799
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1800..2003
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1954..1974
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2005..2023
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2026..2044
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2054..2078
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2151..2160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2160..2183
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2196..2222
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2238..2262
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3976..4000)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4019..4518
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4054..4077)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4451..4470
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4539..5138
                     /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(4539..4558)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5035..5055
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    5724..5740
                     /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(5748..5754,5755..5772,5773..5778))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5793..5814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5931..5948)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6102..6690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6182..6201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6861..7652,7653..7721))
                     /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     7484..7503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7722..7826)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7894..7912
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7950..7972)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8072..8091
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8285..8307
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8435..8454
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatccttg acattgatta ttgactagtt
     1441 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1501 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1561 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1621 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1681 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1741 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1801 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1861 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1921 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1981 gggaggtcta tataagcaga gctctcccta tcagtgatag agatctccct atcagtgata
     2041 gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     2101 gttttgacct ccatagaaga caccgggacc gatccagcct ccggggatct gccaccatgg
     2161 actacaagga tgacgatgac aagctcgatg gaggataccc atacgatgtt ccagattacg
     2221 ctgctcgagg ttatcaaaca agtttgtaca aaaaagttgg catgactata ctccccaaaa
     2281 agaagccgcc gcctcccgac gccgaccccg ccaacgagcc gccgccgccc gggccgatgc
     2341 ccccggcgcc gcggcgcggc ggaggtgtgg gcgtgggcgg cggcggcacg ggcgtgggcg
     2401 gcggcgatcg cgaccgtgac tccggcgtcg tgggggcccg tccgcgagct tcgccaccgc
     2461 ctcaaggccc gctaccagga ccgccgggcg ctcttcatcg ctgggcgctg gccgtgccgc
     2521 ctggtgcagt ggcgggtccc cggccacaac aggcttctcc acctccttgc gggggcccag
     2581 gtggtcccgg cggcggtccc ggcgacgcgc tgggcgcagc ggcggcgggt gtgggtgccg
     2641 cgggcgtggt ggtgggtgtg ggtggtgccg taggcgtggg cggctgctgc tccgggcctg
     2701 ggcacagcaa gcggcgacgt caagctcccg gggttggcgc ggttggcggg ggcagtcccg
     2761 agcgtgagga ggtcggcgca ggctacaaca gtgaggacga gtatgaggcg gctgcagcac
     2821 gcatcgaggc tatggaccct gccactgtcg agcagcagga gcattggttt gaaaaggccc
     2881 tacgagacaa gaagggcttc atcatcaagc agatgaagga ggatggcgcc tgtctcttcc
     2941 gggctgtagc tgaccaggtg tatggagacc aggacatgca tgaggttgtg cgaaagcatt
     3001 gcatggacta tctgatgaag aatgccgact acttctccaa ctatgtcaca gaggacttta
     3061 ccacctacat taacaggaag cggaaaaaca attgccatgg caaccacatt gagatgcagg
     3121 ccatggcaga gatgtacaac cgtcctgtgg aggtgtacca gtacagcaca ggtacttctg
     3181 cagtggaacc catcaacaca ttccatggga tacatcaaaa cgaggacgaa cccattcgtg
     3241 ttagctacca tcggaatatc cactataatt cagtggtgaa tcctaacaag gccaccattg
     3301 gtgtggggct gggcctgcca tcattcaaac cagggtttgc agagcagtct ctgatgaaga
     3361 atgccataaa aacatcggag gagtcatgga ttgaacagca gatgctagaa gacaagaaac
     3421 gggccacaga ctgggaggcc acaaatgaag ccatcgagga gcaggtggct cgggaatcct
     3481 acctgcagtg gttgcgggat caggagaaac aggctcgcca ggtccgaggc cccagccagc
     3541 cccggaaagc cagcgccaca tgcagttcgg ccacagcagc agcctccagt ggcctggagg
     3601 agtggactag ccggtccccg cggcagcgga gttcagcctc gtcacctgag caccctgagc
     3661 tgcatgctga attgggcatg aagccccctt ccccaggcac tgttttagct cttgccaaac
     3721 ctccttcgcc ctgtgcgcca ggtacaagca gtcagttctc ggcaggggcc gaccgggcaa
     3781 cttcccccct tgtgtccctc taccctgctt tggagtgccg ggccctcatt cagcagatgt
     3841 ccccctctgc ctttggtctg aatgactggg atgatgatga gatcctagct tcggtgctgg
     3901 cagtgtccca acaggaatac ctagacagta tgaagaaaaa caaagtgcac agagacccgc
     3961 ccccagacaa gagttgacca actttcttgt acaaagtggt tcgataacga attctaccgg
     4021 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     4081 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     4141 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     4201 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     4261 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     4321 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     4381 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     4441 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     4501 tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac aagcccacgg
     4561 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4621 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4681 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4741 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4801 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4861 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4921 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4981 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5041 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5101 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc
     5161 gcccgaccga aaggagcgca cgaccccatg catcgataaa ataaaagatt ttatttagtc
     5221 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gagaaccatc
     5281 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     5341 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     5401 cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt
     5461 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     5521 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa
     5581 gttggagaac aacattctga gggtaggagt cgaatattaa gtaatcctga ctcaattagc
     5641 cactgttttg aatccacata ctccaatact cctgaaatag ttcattatgg acagcgcaga
     5701 aagagctggg gagaattgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5761 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5821 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5881 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5941 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6001 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6061 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6121 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6181 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6241 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6301 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6361 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6421 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6481 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6541 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6601 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6661 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6721 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6781 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6841 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6901 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6961 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7021 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7081 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7141 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7201 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7261 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7321 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7381 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7441 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7501 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7561 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7621 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7681 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7741 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7801 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7861 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7921 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     7981 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     8041 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     8101 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     8161 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     8221 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     8281 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
     8341 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     8401 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     8461 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
     8521 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     8581 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     8641 agaaaaaggg ggg
//
