LOCUS       sequence-190503-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression construct to express HA-tagged CDC34.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   Feng Q, Jagannathan S, Bradley RK
  TITLE     The RNA Surveillance Factor UPF1 Represses Myogenesis via Its E3 
            Ubiquitin Ligase Activity.
  JOURNAL   Mol Cell. 2017 Jul 20;67(2):239-251.e6. doi: 
            10.1016/j.molcel.2017.05.034. Epub 2017 Jun 29.
  PUBMED    28669802
REFERENCE   2  (bases 1 to 6944)
  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: 
            "10.1016/j.molcel.2017.05.034"; journalName: "Mol Cell"; date: 
            "2017-07-20- 20"; volume: "67"; issue: "2"; pages: "239-251.e6"
COMMENT     Sequence Label: pUB6-CDC34-HA (pRKB261)
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..1229
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1066..1085
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      1461..1470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2145..2156
                     /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)"
     CDS             2175..2201
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2809..2850
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2860..2877
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2900..2917)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2906..3130
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3176..3604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3263..3282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3473..3494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3613..3633)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3618..3947
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3798..3933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3860..3879
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3995..4042
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4061..4459
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4617..4738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4654..4673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4708..4727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4787..4803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4787..4803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4800..4822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4811..4827
                     /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(4835..4841,4842..4859,4860..4865))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4880..4901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5018..5035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5189..5777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5269..5288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5948..6739,6740..6808))
                     /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     6571..6590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6809..6913)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
       61 cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc
      121 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
      181 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
      241 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
      301 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
      361 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
      421 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
      481 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
      541 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
      601 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
      661 caaggtcttg aggccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggg
      721 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
      781 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
      841 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
      901 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
      961 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     1021 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     1081 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     1141 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
     1201 tctggccgtt tttggctttt ttgttagacg aagcttggta ccgagctcgg atccactagt
     1261 ccagtgtggt ggaattcggc cattacggcc ggggagagct gctggagcgc tcggggtccc
     1321 cgggcggcgg cggcggcgca gaggaggagg caggcggcgg ccccggtggc tcccccccgg
     1381 acggtgcgcg gcccggcccg tctcgcgaac tcgcggtggt cgcgcggccc cgcgctgctc
     1441 cgaccccggg cccctccgcc gccgccatgg ctcggccgct agtgcccagc tcgcagaagg
     1501 cgctgctgct ggagctcaag gggctgcagg aagagccggt cgagggattc cgcgtgacac
     1561 tggtggacga gggcgatcta tacaactggg aggtggccat cttcgggccc cccaacacct
     1621 actacgaggg cggctacttc aaggcgcgcc tcaagttccc catcgactac ccatactctc
     1681 caccagcctt tcggttcctg accaagatgt ggcaccctaa catctacgag acgggggacg
     1741 tgtgtatctc catcctccac ccgccggtgg acgaccccca gagcggggag ctgccctcag
     1801 agaggtggaa ccccacgcag aacgtcagga ccattctcct gagtgtgatc tccctcctga
     1861 acgagcccaa caccttctcg cccgcaaacg tggacgcctc cgtgatgtac aggaagtgga
     1921 aagagagcaa ggggaaggat cgggagtaca cagacatcat ccggaagcag gtcctgggga
     1981 ccaaggtgga cgcggagcgt gacggcgtga aggtgcccac cacgctggcc gagtactgcg
     2041 tgaagaccaa ggcgccggcg cccgacgagg gctcagacct cttctacgac gactactacg
     2101 aggacggcga ggtggaggag gaggccgaca gctgcttcgg ggacgatgag gatgactctg
     2161 gcacggagga gtcctaccca tacgatgttc cagattacgc ttgacaccac cagaataaac
     2221 ttgccgagtt tacctcacta gggccggacc cgtggctcct tagacgacag actacctcac
     2281 ggaggttttg tgctggtccc cgtctcctct ggttgtttcg ttttggcttt ttctccctcc
     2341 ccatgtctgt tctgggtttt cacgtgcttc agagaagagg ggctgcccca ccgccactca
     2401 cgtcactcgg ggctcggtgg acgggcccag ggtgggagcg gccggcccac ctgtcccctc
     2461 gggaggggag ctgagcccga cttctaccgg ggtcccccag cttccggact ggccgcaccc
     2521 cggaggagcc acgggggcgc tgctgggaac gtgggcgggg ggccgtttcc tgacactacc
     2581 agcctgggag gcccaggtgt agcggtccga ggggcccggt cctgcctgtc agctccaggt
     2641 cctggagcca cgtccagcac agagtggacg gattcaccgt ggccgactct tttccctgct
     2701 ttggtttgtt tgaaatctaa ataaaactac tttacgaaaa aaaaaaaaaa aaaaaaaaaa
     2761 aaaaacatgt cggccgcctc ggccctcgag tctagagggc ccttcgaagg taagcctatc
     2821 cctaaccctc tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga
     2881 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2941 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3001 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3061 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3121 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3181 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3241 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3301 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3361 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3421 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3481 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3541 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3601 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3781 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3841 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3901 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3961 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     4021 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc ctttgtctca
     4081 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     4141 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     4201 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     4261 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     4321 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     4381 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     4441 ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     4501 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4561 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4741 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4801 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4981 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6241 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6301 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6361 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6421 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6481 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6541 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6601 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6661 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6721 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6781 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6841 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6901 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
