LOCUS       sequence-153740-        4295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector encoding shRNA to UBR5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4295)
  AUTHORS   Shearer RF, Frikstad KM, McKenna J, McCloy RA, Deng N, Burgess A, 
            Stokke T, Patzke S, Saunders DN
  TITLE     The E3 ubiquitin ligase UBR5 regulates centriolar satellite 
            stability and primary cilia.
  JOURNAL   Mol Biol Cell. 2018 May 9:mbcE17040248. doi: 
            10.1091/mbc.E17-04-0248.
  PUBMED    29742019
REFERENCE   2  (bases 1 to 4295)
  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: "Mol Biol 
            Cell. 2018 May 9:mbcE17040248. doi: 10.1091/mbc.E17-04-0248."
COMMENT     Sequence Label: pEN_TmiRc3_shURB5
FEATURES             Location/Qualifiers
     source          1..4295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..48
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    join(205..223,224..246,247..265,266..288,289..307,308..330,
                     331..349,350..372,373..391,392..414,415..433,434..456,
                     457..475)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        508..546
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..567
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     ncRNA           709..830
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           858..872
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           900..1026
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(1065..1164)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1276..1295)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1354..1373)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1947..2480)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2669..2674,2675..2691,2692..2697))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3354..3942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3843..3862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4107..4193
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4285..4295,1..17)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
       61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
      181 tcagtcgact ggatccttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      241 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      301 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      361 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
      421 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
      481 gaaagtcgag ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc
      541 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      601 catagaagac accgggaccg atccagcctc cgcggccccg aactaggcct aggcgtctga
      661 tcactagtga ctctagtcct agtcgactag ggataacagg gtaattgttt gaatgaggct
      721 tcagtacttt acagaatcgt tgcctgcaca tcttggaaac acttgctggg attacttctt
      781 caggttaacc caacagaagg ctcgagaagg tatattgctg ttgacagtga gcgcgcagtg
      841 aatgtagatt ccaaatagtg aagccacaga tgtatttgga atctacattc actgcttgcc
      901 tactgcctcg gaattcaagg ggctacttta ggagcaatta tcttgtttac taaaactgaa
      961 taccttgcta tctctttgat acatttttac aaagctgaat taaaatggta taaattaaat
     1021 cacttttttc aattggaaga ctaatgcgtt ttcgagatat ctagacccag ctttcttgta
     1081 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1141 cagtcaaaat aaaatcatta tttgccatcc agctgcagct ctggcccgtg tctcaaaatc
     1201 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     1261 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     1321 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1381 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1441 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1501 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1561 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     1621 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtc cctgcgccgg ttgcattcga
     1681 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     1741 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     1801 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     1861 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaggccggg
     1921 aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc
     1981 atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa
     2041 tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg
     2101 atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata
     2161 tagatctcac tacgcggctg ctcaaacctg ggcagaacgt aagccgcgag agcgccaaca
     2221 accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg
     2281 aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg
     2341 aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga
     2401 atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca
     2461 tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     2521 acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc
     2581 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     2641 gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg
     2701 ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga
     2761 agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg
     2821 tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctg
     2881 gcttcaggag atcggaagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga
     2941 tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc
     3001 tagctatagt tctagtggtt ggctactaat aggttgtatt gatgttggac gagtcggaat
     3061 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3121 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3181 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3241 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3301 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3361 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3421 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3481 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3541 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3601 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3661 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3721 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3781 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3841 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3901 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3961 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4021 cctgattctg tggataaccg tattaccgct agcatggatc tcggggacgt ctaactacta
     4081 agcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg
     4141 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     4201 gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     4261 aaactgccag gcatcaaact aagcagaagg ccatc
//
