LOCUS       sequence-370238-        8933 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 8933)
  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 8933)
  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-USP5
FEATURES             Location/Qualifiers
     source          1..8933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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             359..873
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(949..1548)
                     /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(1032..1052)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1529..1548
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    1560..2112
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1562..2129
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1722..1741)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1951..1968
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    2142..2166
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    3878..3889
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(4672..4696)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4712..4738)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4751..4774)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4774..4783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4796..5212)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4812..4828)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4844..4866)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5279..5620
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             5684..6200
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(6400..6419)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6547..6569)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6763..6782)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6882..6904
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6942..6960)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7028..7132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7133..7201,7202..7993)
                     /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(7351..7370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8164..8752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8653..8672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8906..8923
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
      241 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
      301 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
      361 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      421 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
      481 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
      541 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      601 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      661 tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      721 cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc
      781 tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc
      841 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa
      901 aatcttttat tttatcgata agcttggctg caggtcgaca tcgatgcatc aggcaccggg
      961 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt
     1021 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg
     1081 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt
     1141 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg
     1201 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc
     1261 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt
     1321 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac
     1381 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac
     1441 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg
     1501 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg ttgtggccat
     1561 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt
     1621 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca
     1681 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc
     1741 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg
     1801 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt
     1861 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa
     1921 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc
     1981 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg
     2041 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc
     2101 ggccagtaac gttagggggg gggggcggaa ttcgttatcg aaccactttg tacaagaaag
     2161 ttgggtagct ggccactctc tggtagaagt agatgtagcc caggtccttg ggcggcttct
     2221 cggaggcaca cactttctgg tcattgtaga tcacccatct gccttctttc ttgatgtggc
     2281 agacgtagtg accacacatg gtagaggtgc ccatgtgact aatgaaggca aagagctgat
     2341 actttccagg accatcccgg actttaggtc ccactggcac agactcagag atggagtcgg
     2401 cagctgagcg gccctctgag atgtccatgg cagcttcagc atccaggtcg tcaatgtgac
     2461 tgaagatcca gtccacagcc cgttctaaac tattgttcgt ggcccgcagc gctttcaagg
     2521 cctggtcccg ggagaagccc atggagacaa tggtggtcac acagtcctca ggaggggggt
     2581 cggctgctgc gcttgtggag cccggcccac tagagccagg caggatgagg gggtttgcaa
     2641 aatctggatc atccatgtgt gacatgaccc agttcatggc ggcctcagcc ccgctgttgc
     2701 ccgtgtagta gacagctttg cggcaggcgt ccatagggaa tcccatctcc accagctgga
     2761 tgatgactga ttcatccagc atgggcgctt tgggctcatc cggagtgacc aggggtgggg
     2821 caatgtctgg cagctcctcc tctccgggct gcagccctgt gcccctcaac tgggagatgt
     2881 cgagctcctc tggcatctcg atggacacat ccagtttctt gggcacccag tctaagccga
     2941 aggtgaactt cttgatctgg atgaccaggt agtcagggaa tgaggcaaat cgtgtggtct
     3001 tgacagctac tgacttggcc tgcagggccg tgctccagaa gtcatcgacc tgctcagggg
     3061 ccccgtaggc ctccaggcaa gagctgaagg gcacctgggc ccgaaccagt tctggcagtg
     3121 ccatcttctc ctcttcggct tgccgcttct tctcctcgta ctccagaagc tcctctttgt
     3181 taagggctgc atccatgggc acaggcagct gcatgatgta gtcaactcgc tgggtgtact
     3241 tcaccttctc tgtggccagg cacttgatct tttcctccac caagaagcgg aacacttcat
     3301 taggattttc agagctccgg caattcctct ccaccatgtt gataaggtga aggaagaact
     3361 cctgggcatc ctgctgccgg ttggtggaga attcagggtg gcccttgccg atgagggcct
     3421 tgaacatccg aggggcaatg ccatcttgaa cttccttctg ttctggcacc cgctccccat
     3481 cgcccgactc cggtactggc ttggaatact ccccggagag aaggccatgg cccagcttgg
     3541 ccacctgggt gctgaaatcc tgggtagggt ccgtcggggc attctggaag atcttctcca
     3601 gcttatccac atacttcctc tggaagtcag ggatgctgaa gagcacctgg accacagagt
     3661 tgaggtagca gctgttaccc aggttccgga tgcctgtgta gccaggccca aacaggggct
     3721 tgagtggcac acctgactcc tggatcagct cccattcacc aatccgctgg ttcatgtcta
     3781 tctccaactc agtcatcgtc ttgtctgtct tctgcatctt cagcatgtcg atgccgaagt
     3841 gggacaggtg ctcagccagg ctggggtcca ggaccatgtc atcctcatca tatgagtaca
     3901 cgtcagctcc atcaggggtg atggtgccca gcttgacagc taacgggtag cctgtctctc
     3961 ggtagtgctc cacagcgtgg ttgttgcccc cactgccatc gaagtagcgt cgcccacaga
     4021 ggatggagcc atcagtcagg ttgagccaca ggttctctct catgtcacac ttggagcact
     4081 tccagccaca gggagggatt cgagcagggt tgtccaactg cttgaggctg aaggcatgct
     4141 tagacacctg ccgtacttcc ccatcccatg cctgcacctc ctgcttgcgg gaggctgagt
     4201 cggccgacag tagggcctcc actgcactgg tcacccgatc tctgacaatg tcaggcagtc
     4261 cccccagtcc atcccgggca atctccaggt aatctggcaa aatgacaatc ttcacatcct
     4321 cgtctaattc aaacttctcc tcgctaaggt caaatccgcc ttcaacacca atagccagcc
     4381 gcgtgggctt cttccggggt gggtctccag tgcctgtagc agggtcctcc tctttcgggc
     4441 gccgggtccg ccggaggtgc aagtagactc gctggccggt cttattgaaa tgtctctcca
     4501 catactgttt cccaaagccc agaaacgtgt tcatacagat gtagaggccc ccctcagact
     4561 ccggcgtgtc gaaggagaag gcgcactcgt ctttgtggac ccggtctcca gccttaggga
     4621 cccggatcgt cggtaatact gacagcagcg cctcctcact cagctccgcc atgccaactt
     4681 ttttgtacaa acttgtttga taacctcgag cagcgtaatc tggaacatcg tatgggtatc
     4741 ctccatcgag cttgtcatcg tcatccttgt agtccatggt ggcagatcta attccggcgc
     4801 ctagagaagg agtgagggct ggataaaggg aggatcgagg cggggtcgaa cgaggaggtt
     4861 caagggggag agacggggcg gatggaggaa gaggaggcgg aggcttaggg tgtacaaagg
     4921 gcttgaccca gggagggggg tcaaaagcca aggcttccca ggtcacgatg taggggacct
     4981 ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat cttaacctgg gtgatgaggt
     5041 ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa ggttggccat tctgcagagc
     5101 agaaggtaac ccaacgtctc ttcttgacat ctaccgactg gttgtgagcg atccgctcga
     5161 catctttcca gtgacctaag gtcaaactta agggagtggt aacagtctgg ccctaatttt
     5221 cagacaaata cagaaacaca gtcagacaga gacaacacag aacgatgctg cagcgctgca
     5281 gcagacaaga cgcgcggctt cggttccaaa ccgaaagcaa aaattcagac ggaggcggga
     5341 actgttttag gttctcgtct cctaccagaa ccacatatcc tgacggggtc ggattccaca
     5401 tcgactccct tcctcaggtc gggccacaaa aacggccccc aaagtccctg ggacgtctcc
     5461 cagggttgcg gccgggtgtt cagaactcgt cagttccacc acgggtccgc cagatacaga
     5521 gctagttagc taactagtac agacgcaggc gcaaacatta gatgccggca caaacacgca
     5581 caaagacaga gacagacacg aaacgaccgc tggccagctt acctcccggc gggggggtcg
     5641 gtggtccctg ggcaggggtc tccaaatctc ggtggaacct ccaaatgaaa gacccccgag
     5701 gtgggcagtc aatcaatctg aggagaccct cccaaggatc agcgagtcca cgattcggat
     5761 gcaaacagca agaggcttta ttgggaatac gggtacccgg gcgacgcagt ctatcggagg
     5821 actggcgcgc cgagtgaggg gttgtgggct cttttattga gctcggggag cagaagcgcg
     5881 cgaacagaag cgagaagcga actgattggt tagttcaaat aaggcacagg gtcatttcag
     5941 gtccttgggg caccctggaa acatctgatg gttctctaga aactgctgag ggcgggaccg
     6001 catctgggga ccatctgttc ttggccctga gccggggcag gaactgctta ccacagatat
     6061 cctgtttggc ccatattctg ctgtctctct gttcctaacc ttgatctgaa cttctctatt
     6121 ctcagttatg tattttccat gccttgcaaa atggcgttac ttaagctagc ttgccaaacc
     6181 tacaggtggg gtctttcatt cccccctttt tctggagact aaataaaatc ttttatttta
     6241 tctatggctc gtactctata ggcttcagct ggtgatattg ttgagtcaaa actagagcct
     6301 ggaccactga tatcctgtct ttaacaaatt ggactaatcg cctctacgcc ggacgcatcg
     6361 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     6421 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     6481 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     6541 cgccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6601 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6661 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     6721 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     6781 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6841 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6901 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     6961 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     7021 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8041 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8101 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8161 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     8221 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     8281 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     8341 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     8401 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8461 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8521 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8581 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8641 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8701 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8761 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8821 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     8881 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
