LOCUS       sequence-153741-        4297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector encoding non-targeting shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4297)
  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 4297)
  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_shNT
FEATURES             Location/Qualifiers
     source          1..4297
                     /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           859..873
                     /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           902..1028
                     /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(1067..1166)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1278..1297)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1356..1375)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1949..2482)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2671..2676,2677..2693,2694..2699))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3356..3944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3845..3864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4109..4195
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4287..4297,1..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 gcgtcgatgc
      841 tctaaggttc tatcaatagt gaagccacag atgtattgat agaaccttag agcatcgctg
      901 cctactgcct cggaattcaa ggggctactt taggagcaat tatcttgttt actaaaactg
      961 aataccttgc tatctctttg atacattttt acaaagctga attaaaatgg tataaattaa
     1021 atcacttttt tcaattggaa gactaatgcg ttttcgagat atctagaccc agctttcttg
     1081 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1141 atcagtcaaa ataaaatcat tatttgccat ccagctgcag ctctggcccg tgtctcaaaa
     1201 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1261 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1321 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1381 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1441 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1501 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1561 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     1621 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg tccctgcgcc ggttgcattc
     1681 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     1741 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1801 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     1861 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaggccg
     1921 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     1981 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     2041 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     2101 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     2161 tatagatctc actacgcggc tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa
     2221 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     2281 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     2341 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     2401 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     2461 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     2521 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     2581 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     2641 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac
     2701 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc
     2761 gaagtcgagg catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat
     2821 cgtcaggcat tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc
     2881 tggcttcagg agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg
     2941 gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac
     3001 tctagctata gttctagtgg ttggctacta ataggttgta ttgatgttgg acgagtcgga
     3061 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3121 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3181 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     3241 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     3301 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3361 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3421 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3481 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3541 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3601 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3661 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3721 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3781 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3841 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3901 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3961 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4021 cccctgattc tgtggataac cgtattaccg ctagcatgga tctcggggac gtctaactac
     4081 taagcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg
     4141 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     4201 gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     4261 ataaactgcc aggcatcaaa ctaagcagaa ggccatc
//
