LOCUS       sequence-129580-        7412 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 7412)
  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 7412)
  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-UCHL5
FEATURES             Location/Qualifiers
     source          1..7412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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             307..821
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(897..1496)
                     /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(980..1000)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1477..1496
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1508..2060
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1510..2077
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1670..1689)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1899..1916
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    2090..2114
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3099..3123)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3139..3165)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3178..3201)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3201..3210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3223..3639)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3239..3255)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3271..3293)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3706..4047
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             4111..4627
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4827..4846)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4974..4996)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5190..5209)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5309..5331
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5369..5387)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5455..5559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5560..5628,5629..6420)
                     /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(5778..5797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6591..7179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7080..7099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7333..7350
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca caattctccc cagctctttc tgcgctgtcc
      181 ataatgaact atttcaggag tattggagta tgtggattca aaacagtggc taattgagtc
      241 aggattactt aatattcgac tcctaccctc agaatgttgt tctccaactt caagaaactg
      301 ttacccatga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      361 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      421 gggcgacgca gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt
      481 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      541 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      601 gaaactgctg agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      661 aggaactgct taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa
      721 ccttgatctg aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt
      781 acttaagcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga
      841 ctaaataaaa tcttttattt tatcgataag cttggctgca ggtcgacatc gatgcatcag
      901 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
      961 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     1021 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     1081 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
     1141 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     1201 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     1261 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     1321 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     1381 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     1441 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt
     1501 gtggccatat tatcatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc
     1561 tagacgtttt tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc
     1621 agatcccata caatggggta ccttctgggc atccttcagc cccttgttga atacgcttga
     1681 ggagagccat ttgactcttt ccacaactat ccaactcaca acgtggcact ggggttgtgc
     1741 cgcctttgca ggtgtatctt atacacgtgg cttttggccg cagaggcacc tgtcgccagg
     1801 tggggggttc cgctgcctgc aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc
     1861 cagaggaact gcttccttca cgacattcaa cagaccttgc attcctttgg cgagagggga
     1921 aagaccccta ggaatgctcg tcaagaagac agggccaggt ttccgggccc tcacattgcc
     1981 aaaagacggc aatatggtgg aaaataacat atagacaaac gcacaccggc cttattccaa
     2041 gcggcttcgg ccagtaacgt tagggggggg gggcggaatt cgttatcgaa ccactttgta
     2101 caagaaagtt gggcatttgg tttcctgagc tttctttgcg ttctgttttt cttttgcctt
     2161 ttctactagt ggtattaact gctggtgttc tgctaaagtc tttaacaatt ccataatgaa
     2221 aggcagataa ttatgcttcc ttctgatatt ctcaatcttg tatcttttta atttctgtac
     2281 ttcttcttca ataagcatct gatttttggc aacttctgac tgaatagcac ttaacatact
     2341 attaccttga tctgtatcca tgggttcctc tgcaagttgt ctttgtaact ctgctatctt
     2401 ctgctcatat atcatttttc tgtcagacac aatggccatt aaattaaatc gaatttcacc
     2461 ttcactgtac ttttgtatcc ttttttctat gacaggcctt actgcactga tccaatcatc
     2521 ttgattgcat gcacctaaat caatcggtcc ttctcttaat ccatctaatt catacagtct
     2581 cccattaaca ggaacataac tgacaaagtg aaaagcatct tcttcttttg ctgatgtctt
     2641 cgtatcaaat tcaaacattt gctgtctggc gaaactgttg tgtacttgtc gaatcacatc
     2701 tgaattgctc agtgccaagc ctttcatagc tgcatcaaaa ctttgtgaaa attctttaaa
     2761 ctctgataat gtctcgccta aatggacatc ctggtgggta cagttcagta acacactcac
     2821 tatggcttga gtagcacaag cattattaat tacctgctta gcaaaaaata tcgtgtcaag
     2881 tcgggagtcc tgaaccacag agcctgctgg ttcttctcct ggctgccact tgaaaagaaa
     2941 aattaaccca tgaactggct ttaatttttc aaaattctca ggctctaaac tccatatttc
     3001 ttctacttgg gctcctcggc aaccgaatcc tttaatgagc tcggtgaaga ccccggggtc
     3061 gctttccatg aggcaccact ccccggcatt gcccgtcatg ccaacttttt tgtacaaact
     3121 tgtttgataa cctcgagcag cgtaatctgg aacatcgtat gggtatcctc catcgagctt
     3181 gtcatcgtca tccttgtagt ccatggtggc agatctaatt ccggcgccta gagaaggagt
     3241 gagggctgga taaagggagg atcgaggcgg ggtcgaacga ggaggttcaa gggggagaga
     3301 cggggcggat ggaggaagag gaggcggagg cttagggtgt acaaagggct tgacccaggg
     3361 aggggggtca aaagccaagg cttcccaggt cacgatgtag gggacctggt ctgggtgtcc
     3421 atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg atgaggtctc ggttaaaggt
     3481 gccgtctcgc ggccatccga cgttaaaggt tggccattct gcagagcaga aggtaaccca
     3541 acgtctcttc ttgacatcta ccgactggtt gtgagcgatc cgctcgacat ctttccagtg
     3601 acctaaggtc aaacttaagg gagtggtaac agtctggccc taattttcag acaaatacag
     3661 aaacacagtc agacagagac aacacagaac gatgctgcag cgctgcagca gacaagacgc
     3721 gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga ggcgggaact gttttaggtt
     3781 ctcgtctcct accagaacca catatcctga cggggtcgga ttccacatcg actcccttcc
     3841 tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga cgtctcccag ggttgcggcc
     3901 gggtgttcag aactcgtcag ttccaccacg ggtccgccag atacagagct agttagctaa
     3961 ctagtacaga cgcaggcgca aacattagat gccggcacaa acacgcacaa agacagagac
     4021 agacacgaaa cgaccgctgg ccagcttacc tcccggcggg ggggtcggtg gtccctgggc
     4081 aggggtctcc aaatctcggt ggaacctcca aatgaaagac ccccgaggtg ggcagtcaat
     4141 caatctgagg agaccctccc aaggatcagc gagtccacga ttcggatgca aacagcaaga
     4201 ggctttattg ggaatacggg tacccgggcg acgcagtcta tcggaggact ggcgcgccga
     4261 gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga
     4321 gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac
     4381 cctggaaaca tctgatggtt ctctagaaac tgctgagggc gggaccgcat ctggggacca
     4441 tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca
     4501 tattctgctg tctctctgtt cctaaccttg atctgaactt ctctattctc agttatgtat
     4561 tttccatgcc ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc
     4621 tttcattccc ccctttttct ggagactaaa taaaatcttt tattttatct atggctcgta
     4681 ctctataggc ttcagctggt gatattgttg agtcaaaact agagcctgga ccactgatat
     4741 cctgtcttta acaaattgga ctaatcgcct ctacgccgga cgcatcgtgg ccggcatcac
     4801 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     4861 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     4921 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgc cgtcgtttta
     4981 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5041 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5101 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5161 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5221 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5281 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5341 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5401 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5461 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5521 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5581 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5641 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5701 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5761 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5821 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5881 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5941 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6001 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6061 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6121 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6181 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6241 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6301 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6361 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6421 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6481 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6541 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6601 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6661 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6721 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6781 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6841 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6901 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6961 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7021 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7081 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7141 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7201 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7261 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7321 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7381 cctctccccg cgcgttggcc gattcattaa tg
//
