LOCUS       sequence-313758-        7499 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 7499)
  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 7499)
  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-EIF3S5
FEATURES             Location/Qualifiers
     source          1..7499
                     /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(1613..1637)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1650..2217
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1667..2219
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1811..1828)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2038..2057
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2231..2830
                     /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(2231..2250)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2727..2747
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2906..3420
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3582..3598
                     /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(3606..3612,3613..3630,3631..3636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3651..3672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3789..3806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3960..4548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4040..4059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4719..5510,5511..5579))
                     /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     5342..5361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5580..5684)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5752..5770
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5808..5830)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5930..5949
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6143..6165
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6293..6312
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6512..7028
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7092..7433
                     /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 atggccacac cggcggtacc agtaagtgct cctccggcca cgccaacccc agtcccggcg
      601 gcggccccag cctcagttcc agcgccaacg ccagcaccgg ctgcggctcc ggttcccgct
      661 gcggctccag cctcatcctc agaccctgcg gcagcagcgg ctgcaactgc ggctcctggc
      721 cagaccccgg cctcagcgca agctccagcg cagaccccag cgcccgctct gcctggtcct
      781 gctcttccag ggcccttccc cggcggccgc gtggtcaggc tgcacccagt cattttggcc
      841 tccattgtgg acagctacga gagacgcaac gagggtgctg cccgagttat cgggaccctg
      901 ttgggaactg tcgacaaaca ctcagtggag gtcaccaatt gcttttcagt gccgcacaat
      961 gagtcagaag atgaagtggc tgttgacatg gaatttgcta agaatatgta tgaactgcat
     1021 aaaaaagttt ctccaaatga gctcatcctg ggctggtacg ctacgggcca tgacatcaca
     1081 gagcactctg tgctgatcca tgagtactac agccgagagg cccccaaccc catccacctc
     1141 actgtggaca caagtctcca gaacggccgc atgagcatca aagcctacgt cagcacttta
     1201 atgggagtcc ctgggaggac catgggagtg atgttcacgc ctctgacagt gaaatacgcg
     1261 tactacgaca ctgaacgcat cggagttgac ctgatcatga agacctgctt tagccccaac
     1321 agagtgattg gactctcaag tgacttgcag caagtaggag gggcatcagc tcgcatccag
     1381 gatgccctga gtacagtgtt gcaatatgca gaggatgtac tgtctggaaa ggtgtcagct
     1441 gacaatactg tgggccgctt cctgatgagc ctggttaacc aagtaccgaa aatagttccc
     1501 gatgactttg agaccatgct caacagcaac atcaatgacc ttttgatggt gacctacctg
     1561 gccaacctca cacagtcaca gattgcactc aatgaaaaac ttgtaaacct gtgcccaact
     1621 ttcttgtaca aagtggttcg ataacgaatt ccgccccccc cccctaacgt tactggccga
     1681 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     1741 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     1801 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     1861 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     1921 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     1981 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     2041 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     2101 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     2161 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     2221 ggccacaacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     2281 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     2341 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     2401 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     2461 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     2521 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     2581 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     2641 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     2701 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     2761 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     2821 cggtgcctga tgcatcgatg tcgacctgca gccaagctta tcgataaaat aaaagatttt
     2881 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     2941 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     3001 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     3061 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     3121 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     3181 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3241 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     3301 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     3361 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     3421 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     3481 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     3541 cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt
     3601 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3661 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3721 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3781 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3841 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3961 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4021 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4141 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4381 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4561 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4621 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4681 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4741 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4801 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4861 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4921 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4981 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5041 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5101 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5161 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5221 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5281 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5341 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5401 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5461 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5521 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5581 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5641 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5701 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5761 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     5821 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     5881 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     5941 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     6001 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6061 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     6121 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg
     6181 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6241 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6301 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6361 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     6421 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     6481 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     6541 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     6601 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
     6661 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
     6721 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     6781 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     6841 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     6901 gactgcgtcg cccgggtacc cgtattccca ataaagcctc ttgctgtttg catccgaatc
     6961 gtggactcgc tgatccttgg gagggtctcc tcagattgat tgactgccca cctcgggggt
     7021 ctttcatttg gaggttccac cgagatttgg agacccctgc ccagggacca ccgacccccc
     7081 cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt
     7141 gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt agctaactag ctctgtatct
     7201 ggcggacccg tggtggaact gacgagttct gaacacccgg ccgcaaccct gggagacgtc
     7261 ccagggactt tgggggccgt ttttgtggcc cgacctgagg aagggagtcg atgtggaatc
     7321 cgaccccgtc aggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
     7381 cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct
     7441 gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattag
//
