LOCUS       sequence-350813-        9005 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 9005)
  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 9005)
  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-OTUD7B
FEATURES             Location/Qualifiers
     source          1..9005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(590..609)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(737..759)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(953..972)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1072..1094
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1132..1150)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1218..1322
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1323..1391,1392..2183)
                     /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(1541..1560)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2354..2942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2843..2862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3096..3113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3230..3251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3266..3271,3272..3289,3290..3296)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3304..3320
                     /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             3482..3996
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(4072..4671)
                     /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(4155..4175)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4652..4671
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    4683..5235
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4685..5252
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4845..4864)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5074..5091
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    5265..5289
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7867..7891)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(7907..7933)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(7946..7969)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      7969..7978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(7991..8407)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(8007..8023)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(8039..8061)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    8474..8815
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             join(8879..9005,1..390)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ctatcggagg actggcgcgc cgagtgaggg gttgtgggct cttttattga gctcggggag
       61 cagaagcgcg cgaacagaag cgagaagcga actgattggt tagttcaaat aaggcacagg
      121 gtcatttcag gtccttgggg caccctggaa acatctgatg gttctctaga aactgctgag
      181 ggcgggaccg catctgggga ccatctgttc ttggccctga gccggggcag gaactgctta
      241 ccacagatat cctgtttggc ccatattctg ctgtctctct gttcctaacc ttgatctgaa
      301 cttctctatt ctcagttatg tattttccat gccttgcaaa atggcgttac ttaagctagc
      361 ttgccaaacc tacaggtggg gtctttcatt cccccctttt tctggagact aaataaaatc
      421 ttttatttta tctatggctc gtactctata ggcttcagct ggtgatattg ttgagtcaaa
      481 actagagcct ggaccactga tatcctgtct ttaacaaatt ggactaatcg cctctacgcc
      541 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
      601 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
      661 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
      721 ccattccttg cgccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
      781 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
      841 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt
      901 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
      961 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1021 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1081 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     1141 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1201 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1261 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1321 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1381 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1441 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1501 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1561 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1621 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1681 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1741 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1801 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     1861 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     1921 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     1981 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2041 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2101 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2161 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2221 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2281 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2341 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2401 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2461 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2521 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2581 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2641 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2701 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2761 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2821 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2881 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2941 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3001 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3061 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3121 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3181 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3241 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     3301 gaattgtgag cggataacaa tttcacaatt ctccccagct ctttctgcgc tgtccataat
     3361 gaactatttc aggagtattg gagtatgtgg attcaaaaca gtggctaatt gagtcaggat
     3421 tacttaatat tcgactccta ccctcagaat gttgttctcc aacttcaaga aactgttacc
     3481 catgaaagac ccccgctgac gggtagtcaa tcactcagag gagaccctcc caaggaacag
     3541 cgagaccaca agtcggatgc aactgcaaga gggtttattg gatacacggg tacccgggcg
     3601 acgcagtcta tcggaggact ggcgcgccga gtgaggggtt gtgggctctt ttattgagct
     3661 cggggagcag aagcgcgcga acagaagcga gaagcgaact gattggttag ttcaaataag
     3721 gcacagggtc atttcaggtc cttggggcac cctggaaaca tctgatggtt ctctagaaac
     3781 tgctgagggc gggaccgcat ctggggacca tctgttcttg gccctgagcc ggggcaggaa
     3841 ctgcttacca cagatatcct gtttggccca tattctgctg tctctctgtt cctaaccttg
     3901 atctgaactt ctctattctc agttatgtat tttccatgcc ttgcaaaatg gcgttactta
     3961 agctagcttg ccaaacctac aggtggggtc tttcattccc ccctttttct ggagactaaa
     4021 taaaatcttt tattttatcg ataagcttgg ctgcaggtcg acatcgatgc atcaggcacc
     4081 gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac
     4141 ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct ccaggaaggc
     4201 gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc tgcccagacc
     4261 cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg
     4321 ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg aaccgctcaa
     4381 ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt
     4441 ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc
     4501 gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc
     4561 gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct
     4621 ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tggttgtggc
     4681 catattatca tcgtgttttt caaaggaaaa ccacgtcccc gtggttcggg gggcctagac
     4741 gtttttttaa cctcgactaa acacatgtaa agcatgtgca ccgaggcccc agatcagatc
     4801 ccatacaatg gggtaccttc tgggcatcct tcagcccctt gttgaatacg cttgaggaga
     4861 gccatttgac tctttccaca actatccaac tcacaacgtg gcactggggt tgtgccgcct
     4921 ttgcaggtgt atcttataca cgtggctttt ggccgcagag gcacctgtcg ccaggtgggg
     4981 ggttccgctg cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa gcttccagag
     5041 gaactgcttc cttcacgaca ttcaacagac cttgcattcc tttggcgaga ggggaaagac
     5101 ccctaggaat gctcgtcaag aagacagggc caggtttccg ggccctcaca ttgccaaaag
     5161 acggcaatat ggtggaaaat aacatataga caaacgcaca ccggccttat tccaagcggc
     5221 ttcggccagt aacgttaggg ggggggggcg gaattcgtta tcgaaccact ttgtacaaga
     5281 aagttgggta tcagaacctg tgcaccagga gctccccatc cggttcccgc tccctcctcc
     5341 tcagttcttc cctgtaacaa caggaacaga agttgtttgt ctcagggtgt ccatagaagc
     5401 tgcagttcgg ttgtttgcat ttggtctgag ttgggggaag gccccggaga cctccagccc
     5461 atccatctgg ctcagggggc tctctgtagc cattgctata ggaatcagcc actcggtagg
     5521 ggggtggtaa caaggcaccc ctgtgaagtc catccttaga gtggctgcct ggctccagag
     5581 aagggatgct gtcctggtgg gggtagggtc gcccaggagg gcactgtctg gggaaggtgg
     5641 catatggtgg taggcccccg acacatggac cccctgccaa ctgcctccgg ggttcctggc
     5701 agtggactcc gcccccagac ggccgaggga tagtaaagtc cccagggtag ccagtggaaa
     5761 atgccattgc cctggactct gctgggggac cggtcggctg ctcttcccta gcatctggct
     5821 ctggcttttt ggctggagga gggccacccc ctattcctcc attcatgatc ttcctctctg
     5881 cctccttctg cttctgttct gccaggaatc tctcctcagc atcagaaagg tagcgctgga
     5941 tcatttcctc ctgatactgg tgacggtgac ccatcttcag ggttccaaca aaaataaact
     6001 tcccctcccc ttgcatggca gtcctcagaa tgctcaggct ctgcatcacc tcctggctat
     6061 acttgctccc tccgttacca acagactcag ctgggggctt ctcagacaca ggcccatccc
     6121 cagctgcctc ctccttgcca cccttccagc tcttcagtga gtttttcttc ttcttctcca
     6181 gtgtctcagt gccgctgctt cctcccaacc ctgtccccac ccctccaggc tttgaaccct
     6241 tgctgtgcat caggcccccc atgttcttct tgagcttgct gcccaaggtt ttgccaaagc
     6301 tgcccagttt gttagccaca gaatctgctc tcttcttgtc cttctcccga tctcgctttg
     6361 acttctcctt ccgccggccg ccctcgttgc tggtggaact gctgccaact gactccttgt
     6421 ctgagtctcc agactcagga gtggaccggg gctcatctcc agctgaggcg gtgggggact
     6481 caggctgggc cagaggagcc tgtgcatcag aggacagtgg gatccacttc acattcatgt
     6541 agctatgcag cagatgcaat ttgacctcta gggacagaat tacactggcc aatcggacat
     6601 tgtcactatc atctttgccc cactcccagc cctttccagg gtccacagca aagtgcaagg
     6661 gcagcagctt atactctgaa tctgtaagtg ggatcacagc ttgttccttg gtattctcct
     6721 tctgctccat ggacacgagt gcagaaaagt gggcctgatc ataggcgagc accagagggg
     6781 agcggtgaca ctggctggct gggacctcca aaggcagata gattcctcca aagggaatag
     6841 gggcaaatgc ttcccctccg gagtccctca gcatggtgtc tgccacgacg actatgggcc
     6901 tcctaagcac atgagcaagg acaaagacgt gaaactcttc aaggctctca tatacaggct
     6961 cctcagaact ctccacccca ccacagttgg ctccattggt acctagatgc attcggggtt
     7021 cacttgaggc aagcttgatc agttcattcc actccttctg ccattcatct tctgtgtata
     7081 ccagccctga ctctttattc tgctgtgtct gctgccacct ccagcgcctt ttcaacgctt
     7141 ccttctcaac tcccttctcc atcagtgcat acaaagcttt ccgcagcatc aagtcccgat
     7201 catggaaacc ccacattcca agggaggctg catgcaggag gcagttccca tctccagtag
     7261 ttgccaaagg aagcagcctc tgagaggtgg gatccacact cacccaccag ttcaaacgcc
     7321 ctgcctgttc caaggcaacc agcatggact gctcaatgag gtctctctct atgaagctgc
     7381 ggaagtcttc attgtataca gtgagatctg gaagctggaa ggcacagatg ggcatttcca
     7441 gggggtgctc attgctcccc ccacccccac cattggagga gacatgggac cgggccaggg
     7501 aaacaatgct ggagctggcg tgggagatgc ccctagacag gcgtttttct tgaacgatgt
     7561 catcctgccg ctggaggatg ggtcggggag ggcgagtagg ctctctgtca gaaaaccctt
     7621 tttcaggggt cctggagcca ccactcccct cactaaagga tgggggtagg tttccagcat
     7681 ggacttgacg tagctgttca aaatcactga gggcggcatt cacatcccaa ttctttcctt
     7741 ctaggagatc tcgcgctagc cctggctctg ctcctgtgga acggacaaaa tctgacagaa
     7801 cagcatccat gtccagggtc atgtgatcct caagtacttt cagccagctg gatgtatggt
     7861 agaacatgcc aacttttttg tacaaacttg tttgataacc tcgagcagcg taatctggaa
     7921 catcgtatgg gtatcctcca tcgagcttgt catcgtcatc cttgtagtcc atggtggcag
     7981 atctaattcc ggcgcctaga gaaggagtga gggctggata aagggaggat cgaggcgggg
     8041 tcgaacgagg aggttcaagg gggagagacg gggcggatgg aggaagagga ggcggaggct
     8101 tagggtgtac aaagggcttg acccagggag gggggtcaaa agccaaggct tcccaggtca
     8161 cgatgtaggg gacctggtct gggtgtccat gcgggccagg tgaaaagacc ttgatcttaa
     8221 cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg ccatccgacg ttaaaggttg
     8281 gccattctgc agagcagaag gtaacccaac gtctcttctt gacatctacc gactggttgt
     8341 gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa acttaaggga gtggtaacag
     8401 tctggcccta attttcagac aaatacagaa acacagtcag acagagacaa cacagaacga
     8461 tgctgcagcg ctgcagcaga caagacgcgc ggcttcggtt ccaaaccgaa agcaaaaatt
     8521 cagacggagg cgggaactgt tttaggttct cgtctcctac cagaaccaca tatcctgacg
     8581 gggtcggatt ccacatcgac tcccttcctc aggtcgggcc acaaaaacgg cccccaaagt
     8641 ccctgggacg tctcccaggg ttgcggccgg gtgttcagaa ctcgtcagtt ccaccacggg
     8701 tccgccagat acagagctag ttagctaact agtacagacg caggcgcaaa cattagatgc
     8761 cggcacaaac acgcacaaag acagagacag acacgaaacg accgctggcc agcttacctc
     8821 ccggcggggg ggtcggtggt ccctgggcag gggtctccaa atctcggtgg aacctccaaa
     8881 tgaaagaccc ccgaggtggg cagtcaatca atctgaggag accctcccaa ggatcagcga
     8941 gtccacgatt cggatgcaaa cagcaagagg ctttattggg aatacgggta cccgggcgac
     9001 gcagt
//
