LOCUS       sequence-129579-        7910 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 7910)
  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 7910)
  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-USP14
FEATURES             Location/Qualifiers
     source          1..7910
                     /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"
     regulatory      1418..1427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1427..1450
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1463..1489
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1505..1529
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3012..3036)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3049..3616
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3066..3618
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3210..3227)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3437..3456
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3630..4229
                     /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(3630..3649)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4126..4146
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4305..4819
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4981..4997
                     /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(5005..5011,5012..5029,5030..5035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5050..5071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5188..5205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5359..5947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5439..5458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6118..6909,6910..6978))
                     /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     6741..6760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6979..7083)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7151..7169
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7207..7229)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7329..7348
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7542..7564
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7692..7711
                     /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 tagatctgcc accatggact acaaggatga
     1441 cgatgacaag ctcgatggag gatacccata cgatgttcca gattacgctg ctcgaggtta
     1501 tcaaacaagt ttgtacaaaa aagttggcat gccgctctac tccgttactg taaaatgggg
     1561 aaaggagaaa tttgaaggtg tagaattgaa tacagatgaa cctccaatgg tattcaaggc
     1621 tcagctgttt gcgttgactg gagtccagcc tgccagacag aaagttatgg tgaaaggagg
     1681 aacgctaaag gatgatgatt ggggaaacat caaaataaaa aatggaatga ctctactaat
     1741 gatggggtca gcagatgctc ttccagaaga accctcagcc aaaactgtct tcgtagaaga
     1801 catgacagaa gaacagttag catctgctat ggagttacca tgtggattga caaaccttgg
     1861 taacacttgt tacatgaatg ccacagttca gtgtattcgt tctgtgcctg aactcaaaga
     1921 tgcccttaaa aggtatgcag gtgccttgag agcttcaggg gaaatggctt cagcgcagta
     1981 tattactgca gcccttagag atttgtttga ttccatggat aaaacttctt ccagtattcc
     2041 acctattatt ctactgcagt ttttgcacat ggctttccca cagtttgccg agaaaggtga
     2101 acaaggacag tatcttcaac aggatgctaa tgaatgttgg atacaaatga tgcgagtatt
     2161 gcaacagaaa ttggaagcaa tagaggatga ttctgttaaa gagacagact cctcatctgc
     2221 atcggcagcg acaccttcta aaaagaaaag tttaatcgat cagttcttcg gtgttgagtt
     2281 tgaaactacc atgaaatgta cagaatctga agaagaagaa gtcaccaaag gaaaggaaaa
     2341 tcaacttcag cttagctgtt ttatcaatca ggaagtcaag tatcttttta caggacttaa
     2401 attgcgactt caggaagaaa tcaccaaaca gtctccaacg ttgcaaagaa atgccttgta
     2461 tatcaaatct tccaagatca gccggctgcc tgcttacttg accattcaga tggttcgatt
     2521 tttttataaa gagaaggaat ctgtgaatgc caaagttctt aaggatgtta aatttcctct
     2581 tatgttggat atgtatgaac tgtgtacacc agaacttcaa gagaaaatgg tgtcttttcg
     2641 atccaaattc aaggatctag aagataaaaa agtgaatcag cagccaaata caagtgacaa
     2701 aaagagtagt ccccagaaag aagttaagta tgaacccttt tcttttgctg atgatattgg
     2761 ctccaataat tgtggatact atgacttaca agcagtacta acacaccagg gaaggtctag
     2821 ttcttcaggt cattatgtat catgggtgaa aaggaaacaa gatgaatgga ttaagtttga
     2881 tgatgacaaa gtcagcatcg taacaccaga agatatctta cggctttctg gtggtggaga
     2941 ctggcatatc gcttacgttc tactctatgg gcctcgcaga gttgaaataa tggaagagga
     3001 aagtgaacag tacccaactt tcttgtacaa agtggttcga taacgaattc cgcccccccc
     3061 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3121 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3181 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3241 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3301 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3361 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3421 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3481 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     3541 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3601 aaaaacacga tgataatatg gccacaacca tgaccgagta caagcccacg gtgcgcctcg
     3661 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3721 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     3781 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     3841 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     3901 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     3961 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4021 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4081 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4141 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4201 ggtgcatgac ccgcaagccc ggtgcctgat gcatcgatgt cgacctgcag ccaagcttat
     4261 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     4321 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     4381 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     4441 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     4501 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     4561 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     4621 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     4681 agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     4741 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     4801 gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac attctgaggg
     4861 taggagtcga atattaagta atcctgactc aattagccac tgttttgaat ccacatactc
     4921 caatactcct gaaatagttc attatggaca gcgcagaaag agctggggag aattgtgaaa
     4981 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5041 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5101 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5161 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5221 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5281 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5341 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5401 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5461 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5521 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5581 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5641 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5701 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5761 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5821 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5881 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5941 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6001 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6061 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6121 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6181 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6241 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6301 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6361 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6421 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6481 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6541 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6601 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6661 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6721 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6781 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6841 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6901 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6961 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7021 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7081 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7141 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     7201 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     7261 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     7321 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     7381 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     7441 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     7501 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     7561 aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     7621 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     7681 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     7741 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata
     7801 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac
     7861 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg
//
