LOCUS       sequence-313505-        7952 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 7952)
  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 7952)
  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-USP30
FEATURES             Location/Qualifiers
     source          1..7952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      430..439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             439..462
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             475..501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    517..541
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2066..2090)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2103..2670
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2120..2672
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2264..2281)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2491..2510
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2684..3283
                     /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(2684..2703)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3180..3200
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3359..3873
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4035..4051
                     /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(4059..4065,4066..4083,4084..4089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4104..4125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4242..4259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4413..5001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4493..4512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5172..5963,5964..6032))
                     /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     5795..5814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6033..6137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6205..6223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6261..6283)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6383..6402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6596..6618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6746..6765
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6965..7481
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7545..7886
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
      421 attagatctg ccaccatgga ctacaaggat gacgatgaca agctcgatgg aggataccca
      481 tacgatgttc cagattacgc tgctcgaggt tatcaaacaa gtttgtacaa aaaagttggc
      541 atgaccgcgg ccgacagggc catccagcgc ttcctgcgga ccggggcggc cgtcagatat
      601 aaagtcatga agaactgggg agttataggt ggaattgctg ctgctcttgc agcaggaata
      661 tatgttattt ggggtcccat tacagaaaga aagaagcgta gaaaagggct tgtgcctggc
      721 cttgttaatt tagggaacac ctgcttcatg aactccctgc tacaaggcct gtctgcctgt
      781 cctgctttca tcaggtggct ggaagagttc acctcccagt actccaggga tcagaaggag
      841 cccccctcac accagtattt atccttaaca ctcttgcacc ttctgaaagc cttgtcctgc
      901 caagaagtta ctgatgatga ggtcttagat gcaagctgct tgttggatgt cttaagaatg
      961 tacagatggc agatctcatc atttgaagaa caggatgctc acgaattatt ccatgtcatt
     1021 acctcgtcat tggaagatga gcgagaccgc cagcctcggg tcacacattt gtttgatgtg
     1081 cattccctgg agcagcagtc agaaataact cccaaacaaa ttacctgccg cacaagaggg
     1141 tcacctcacc ccacatccaa tcactggaag tctcaacatc cttttcatgg aagactcact
     1201 agtaatatgg tctgcaaaca ctgtgaacac cagagtcctg ttcgatttga tacctttgat
     1261 agcctttcac taagtattcc agccgccaca tggggtcacc cattgaccct ggaccactgc
     1321 cttcaccact tcatctcatc agaatcagtg cgggatgttg tgtgtgacaa ctgtacaaag
     1381 attgaagcca agggaacgtt gaacggggaa aaggtggaac accagaggac cacttttgtt
     1441 aaacagttaa aactagggaa gctccctcag tgtctctgca tccacctaca gcggctgagc
     1501 tggtccagcc acggcacgcc tctgaagcgg catgagcacg tgcagttcaa tgagttcctg
     1561 atgatggaca tttacaagta ccacctcctt ggacataaac ctagtcaaca caaccctaaa
     1621 ctgaacaaga acccagggcc tacactggag ctgcaggatg ggccgggagc ccccacacca
     1681 gttctgaatc agccaggggc ccccaaaaca cagattttta tgaatggcgc ctgctcccca
     1741 tctttattgc caacgctgtc agcgccgatg cccttccctc tcccagttgt tcccgactac
     1801 agctcctcca catacctctt ccggctgatg gcagttgtcg tccaccatgg agacatgcac
     1861 tctggacact ttgtcactta ccgacggtcc ccaccttctg ccaggaaccc tctctcaact
     1921 agcaatcagt ggctgtgggt ctccgatgac actgtccgca aggccagcct gcaggaggtc
     1981 ctgtcctcca gcgcctacct gctgttctac gagcgcgtcc tttccaggat gcagcaccag
     2041 agccaggagt gcaagtctga agaatgccca actttcttgt acaaagtggt tcgataacga
     2101 attccgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2161 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2221 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2281 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     2341 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2401 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2461 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2521 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2581 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2641 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatgaccg agtacaagcc
     2701 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     2761 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     2821 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     2881 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     2941 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     3001 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     3061 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     3121 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     3181 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     3241 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgatgcatcg atgtcgacct
     3301 gcagccaagc ttatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     3361 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     3421 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag
     3481 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3541 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc
     3601 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     3661 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     3721 tcactcggcg cgccagtcct ccgatagact gcgtcgcccg ggtacccgtg tatccaataa
     3781 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     3841 gtgattgact acccgtcagc gggggtcttt catgggtaac agtttcttga agttggagaa
     3901 caacattctg agggtaggag tcgaatatta agtaatcctg actcaattag ccactgtttt
     3961 gaatccacat actccaatac tcctgaaata gttcattatg gacagcgcag aaagagctgg
     4021 ggagaattgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6181 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6241 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6301 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6361 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     6421 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     6481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     6601 tcacgacgtt gtaaaacgac ggcgcaagga atggtgcatg caaggagatg gcgcccaaca
     6661 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6721 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6781 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggcgatta gtccaatttg
     6841 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     6901 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     6961 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     7021 aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag gaacagagag
     7081 acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     7141 ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga gaaccatcag
     7201 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     7261 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     7321 caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt acccgtattc
     7381 ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct tgggagggtc
     7441 tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc caccgagatt
     7501 tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg gccagcggtc
     7561 gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt ttgcgcctgc
     7621 gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     7681 tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg
     7741 gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg
     7801 taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga
     7861 accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc
     7921 tgactgtgtt tctgtatttg tctgaaaatt ag
//
