LOCUS       sequence-287545-        8942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged mouse DNA topoisomerase IIbeta (full length 
            cDNA, in pCS2+ backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8942)
  AUTHORS   Ban Y, Ho CW, Lin RK, Lyu YL, Liu LF
  TITLE     Activation of a novel ubiquitin-independent proteasome pathway when 
            RNA polymerase II encounters a protein roadblock.
  JOURNAL   Mol Cell Biol. 2013 Oct;33(20):4008-16. doi: 10.1128/MCB.00403-13. 
            Epub 2013 Aug 12.
  PUBMED    23938298
REFERENCE   2  (bases 1 to 8942)
  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"; doi: 
            "10.1128/MCB.00403-13"; journalName: "Mol Cell Biol"; date: 
            "2013-10"; volume: "33"; issue: "20"; pages: "4008-16"
COMMENT     Sequence Label: pYLL201
FEATURES             Location/Qualifiers
     source          1..8942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..984
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1019..1037
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1019..1036
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1086..1112
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    5978..6112
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6002..6021)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6056..6075
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6225..6245)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6225..6243)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6264..6280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6264..6280)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6277..6299)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6288..6304
                     /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(6312..6318,6319..6336,6337..6342))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6357..6378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6495..6512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6666..7254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6746..6765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7425..8216,8217..8285))
                     /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     8048..8067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8286..8390)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8416..8871)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8553..8574)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8765..8784
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
       61 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
      121 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
      181 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
      241 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
      301 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
      361 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
      421 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
      481 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
      541 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
      601 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
      661 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
      721 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
      781 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
      841 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
      901 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
      961 ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat
     1021 ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca tcgatagcca
     1081 ccatgtaccc atacgacgtc ccagactacg ctgccaagtc cagcctcgcg ggctctgacg
     1141 gggcgctgac ctgggtgaac aatgctacaa aaaaggaaga gttggagaca gcaaacaaaa
     1201 atgactctac taagaagttg tctgttgaga gagtgtacca gaagaagacc caactggagc
     1261 acattctcct tcgtcctgat acatacattg gatcagtgga gccactgaca cagctaatgt
     1321 gggtgtatga tgaagatgta gggatgaact gcagagaggt tacctttgtt ccaggcttat
     1381 acaagatctt tgatgaaatt ttggttaatg ctgctgacaa caagcagagg gacaagaaca
     1441 tgacctgtat taaggtttct attgatcctg aatctaatat tataagcatt tggaataatg
     1501 ggaaaggcat tccagtagta gaacacaaag tagagaaagt ttatgttcct gctttaattt
     1561 ttggacagct tttaacatcg agtaattatg acgatgatga gaaaaaagtt acaggaggcc
     1621 gtaatggtta tggtgcaaag ctttgtaata tttttagtac aaagtttaca gtagaaacgg
     1681 cttgcaaaga atacaaacat agttttaaac agacatggat gaataatatg atgaagactt
     1741 ctgaagccaa aattaaacat tttgatggtg aagattatac atgcataaca ttccagccag
     1801 atctatccaa atttaagatg gaaaaacttg acaaggacat tgtggccctc atgaccagaa
     1861 gagtatatga tttagctggg tcctgtaaag gagtcaaggt catgtttaat gggaagaagt
     1921 tgcctgtaaa tggatttcgc agctatgtag acctttatgt gaaagacaaa ttggatgaaa
     1981 ctggggtggc tctcaaagtt attcatgaac ttgcaaatga aagatgggat gtttgtctaa
     2041 ccttgagtga aaaaggtttc cagcagatca gctttgtgaa tagtattgca actacaaaag
     2101 gtgggcggca tgtggattat gtagtagatc aagttgttag taaactgatt gaagtagtta
     2161 agaaaaagaa caaagctggt gtatcagtga aaccatttca ggttaaaaac catatatggg
     2221 tttttattaa ttgtcttatt gaaaatccaa cctttgattc tcaaactaag gaaaacatga
     2281 ctctgcagcc caaaagtttt gggtccaaat gccaactgtc agaaaaattt tttaaagcag
     2341 cctctaattg tggcattgta gaaagcatat tgaactgggt gaaatttaag gctcagactc
     2401 aactgaataa gaagtgctca tcagtaaagt acagtaaaat caaaggcatt cccaaactgg
     2461 atgatgccaa tgatgctggt ggcaaacatt ccctggagtg tacattaata ttaacagagg
     2521 gggactccgc caaatctcta gctgtatctg gattaggtgt gattggtagg gacagatatg
     2581 gagtcttccc actcaggggt aaaattctca atgtacgaga agcctctcat aaacagatta
     2641 tggaaaatgc agagataaat aatattatta aaatagttgg tttgcaatac aagaaaagtt
     2701 atgatgatgc agaatctctg aaaactttac gttatggaaa gattatgatt atgactgatc
     2761 aggatcaaga tggttctcac atcaaaggcc tgctaatcaa ttttattcat cacaattggc
     2821 catcactttt gaagcatggt tttcttgaag aattcattac tcccattgta aaggcaagca
     2881 aaaataagca ggaactttcc ttttatagta ttcctgaatt tgatgagtgg aaaaaacata
     2941 tagaaaacca gaaagcctgg aaaataaagt actacaaagg attaggtacc agtaccgcta
     3001 aagaagcaaa ggaatatttt gctgatatgg aaaggcatcg cattttgttt agatatgctg
     3061 gacctgaaga tgatgctgcc atcaccttgg catttagtaa gaagaagatt gatgacagaa
     3121 aagagtggtt aaccaacttt atggaggatc ggagacagcg taggttacat ggcctaccag
     3181 agcaattttt atatggtaca gcaacaaagc atttgacata caatgatttc atcaacaagg
     3241 aattgattct cttctcaaac tcggacaatg agagatctat cccatctctt gtggatggct
     3301 tcaagccagg tcaacgaaaa gttttgttta cctgtttcaa gaggaatgat aaacgtgaag
     3361 tgaaagtagc gcagctggct ggatctgttg ctgagatgtc tgcttaccac catggagaac
     3421 aagcattgat gatgactatt gtgaatttgg ctcagaattt tgtgggcagt aataacatta
     3481 acttgcttca gcccattggt cagtttggaa ctcggcttca tggtggcaaa gatgctgcaa
     3541 gccctcgtta tattttcaca atgttgagct ctctggcaag acttcttttt ccagctgtgg
     3601 atgataactt gcttaagttc ctttatgatg ataatcaacg tgtagagcca gagtggtata
     3661 tccctataat ccccatggtt ttgattaatg gtgctgaggg tattgggact ggatgggctt
     3721 gtaaactgcc caactatgat gctagagaaa ttgtgaacaa tgtcaggcgg atgctagaag
     3781 gtttggatcc tcatcccatg cttccaaact ataagaactt caaaggcacc atccaggaac
     3841 ttggtcagaa ccagtatgct gttagtggtg agatatttgt ggtggacaga aacacagtag
     3901 agattacaga acttccagtt agaacatgga cacaggtata caaggagcag gtcctagaac
     3961 ctatgcttaa tggaacagat aaaacaccgg cactgatttc tgattacaaa gaatatcata
     4021 ctgacacaac tgtgaagttt gtggtgaaaa tgacagaaga gaaactagcg caagcagaag
     4081 ctgctggcct gcataaagtt tttaaacttc aaactactct tacttgcaat tccatggtac
     4141 tctttgatca tatgggatgt ctgaagaagt atgaaactgt gcaagacatt ttgaaagaat
     4201 tctttgattt acggttaagt tattatggtt tacggaagga atggcttgta ggaatgctgg
     4261 gagcagaatc tacaaagctt aataatcaag cccgtttcat tttagagaag atacagggga
     4321 aaattactat agagaatagg tcaaagaaag atttgattca aatgttagtc cagagaggtt
     4381 atgaatctga cccagtgaaa gcgtggaaag aagcacaaga aaaggctgca gaagaggaag
     4441 attcacaaaa ccaacatgat gacagttcct ctgattcagg aactccttca ggccctgatt
     4501 ttaattatat tttaaatatg tctctatggt ctcttactaa agaaaaagtt gaagaactca
     4561 ttaaacagag agatacgaaa gggcgagagg ttaatgatct caagaggaag tctccttcag
     4621 atctttggaa agaagactta gcagcatttg ttgaagaatt ggataaagtg gaagcacaag
     4681 agcgggaaga tattctggca ggaatgtctg ggaaagcaat aaaaggtaaa gtcggcaaac
     4741 ccaaagtaaa gaaacttcag ctggaggaga caatgccctc gccttacggg agaagaattg
     4801 ttccagagat tactgctatg aaggcagatg ccagcagaaa attgctgaag aagaaaaagg
     4861 gtgatcctga taccacagta gtgaaagtag aatttgatga agaatttagt ggaacaccag
     4921 cagaaggtac aggagaggag acattgaccc catctgctcc tgtaaataaa ggccccaaac
     4981 ccaaaagaga gaagaaggag cctggtacca gggttagaaa aacacctaca tcaactggta
     5041 aaactaatgc gaagaaagtg aagaaacgga atccttggtc agatgatgag tccaaatcag
     5101 agagtgactt ggaagaagca gaacctgtgg ttatcccaag agattctttg cttaggagag
     5161 cagctgctga aagacctaag tacacttttg atttctcaga agaggaggat gatgatgctg
     5221 ctgctgctga tgacagtaat gatttagaag aactgaaagt taaagcatct cccataacaa
     5281 atgatgggga agatgaattt gttccttcag atggcttaga taaagatgag tatgcttttt
     5341 cttcaggcaa atccaaagct actccagaaa aatcttcaaa tgacaaaaaa agtcaggatt
     5401 ttggaaatct tttttcattt ccttcatatt ctcagaagtc agaagatgat tcagctaaat
     5461 ttgacagtaa tgaagaagat accgcttctg tttttgctcc atcatttggt ctgaagcaaa
     5521 cagataaact tccaagtaaa acagttgctg ctaaaaaggg taaaccacct tcagatacag
     5581 cccctaaagc taagagagcc cctaaacaga agaagatagt agagactata aactctgact
     5641 cagactctga atttggcatt ccaaagaaga ctacaacacc aaaaggtaaa ggccgagggg
     5701 caaagaaaag gaaagcatca ggctcagaaa atgaaggtga ttataaccct ggaaggaaac
     5761 catctaaaac agccagcaag aaaccgaaga aaacatcttt tgatcaggat tcagatgtag
     5821 acatcttccc ctcagacttc acttctgaac caccggctct tccacggact ggtcgggcta
     5881 ggaaagaagt aaaatatttt gcagagtctg atgaagaaga agatgttgat tttgcaatgt
     5941 ttaattaatc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     6001 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6061 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     6121 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     6181 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     6241 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6301 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     6361 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6421 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6481 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7381 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7441 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7501 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7561 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7621 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7681 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7741 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7801 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7861 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7921 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7981 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8041 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8101 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8161 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8221 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8281 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8341 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8401 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     8461 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     8521 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     8581 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     8641 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     8701 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     8761 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     8821 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     8881 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     8941 ag
//
