LOCUS       sequence-161213-        4238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5'entry vector containing human ubiquitin C promoter followed by 
            intronically-expressed eSIBR cassette for ubiquitous expression, 
            RNAi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4238)
  AUTHORS   Fowler DK, Stewart S, Seredick S, Eisen JS, Stankunas K, Washbourne 
            P
  TITLE     A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate 
            Expression Constructs with Diverse Research Applications.
  JOURNAL   PLoS One. 2016 Aug 8;11(8):e0159277. doi: 
            10.1371/journal.pone.0159277. eCollection 2016.
  PUBMED    27500400
REFERENCE   2  (bases 1 to 4238)
  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: "PLoS One.";
            date: "2016-08-8"; volume: "11(8):e0159277. doi"; pages: " 
            10.1371/journal.pone.0159277. eCollection 2016"
COMMENT     Sequence Label: p5E-Ui4-eSIBR
FEATURES             Location/Qualifiers
     source          1..4238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(143..162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(798..1607)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1521..1540
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1599..1618
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1720..1736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1720..1736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1741..1760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1741..1759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1775..1870
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1893..2292
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2862..2881
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     3765..3784
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    3814..3937
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4130..4147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
       61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat
      721 tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
      781 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
      841 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
      901 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
      961 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1021 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     1081 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1141 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     1201 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1261 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg
     1321 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1381 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1441 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     1501 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1561 ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac accccttgta
     1621 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     1681 atgtaacatc agagattttg agacacgggc cagagctgcc aggaaacagc tatgaccatg
     1741 taatacgact cactataggg gatcatggag ctccaaataa tgattttatt ttgactgata
     1801 gtgacctgtt cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtat
     1861 agaaaagttg ggccggccct tcggcgcaaa ctggcctccg cgccgggttt tggcgcctcc
     1921 cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg
     1981 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     2041 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     2101 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     2161 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     2221 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     2281 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     2341 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     2401 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     2461 tgaatagaac cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc
     2521 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     2581 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     2641 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     2701 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     2761 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     2821 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     2881 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     2941 atcatttggg tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg
     3001 gccgtttttg gcttttttgt tagacgaagc taaggcgcgc ctgagaactt cagggtgagt
     3061 ttggggaccc ttgattgttc tttctttttc gctattgtaa aattcatgtt atatggaggg
     3121 ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc ccttgtatca ccatgcatgg
     3181 accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat tgtctcctct
     3241 tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt tgtaacgaat
     3301 ttttaaattc acttttgttt atttgtcaga ttgtaagatc ccatcgattc gaattcaaag
     3361 tcgactggag gcttgctttg ggctgtatgc tgttgtcttc aagatctgga agacacagga
     3421 cacaaggccc tttatcagca ctcacatgga acaaatggcc accgtgggag gatgacaact
     3481 cgagcctcta gacaatcagg gtatattata ttgtacttca gcacagtttt agagaacaat
     3541 tgttataatt aaatgataag gtagaatatt tctgcatata aattctggct ggcgtggaaa
     3601 tattcttatt ggtagaaaca actacatcct ggtcatcatc ctgcctttct ctttatggtt
     3661 acaatgatat acactgtttg agatgaggat aaaatactct gagtccaaac cgggcccctc
     3721 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     3781 tgtgctgtct catcaagctt aaggatccgc gcccaagttt gtacaaaaaa gttgaacgag
     3841 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     3901 cataatactg taaaacacaa catatgcagt cactatgaac caactactta gatggtatta
     3961 gtgacctgta gaattatcgc ggccgccctg cagctctagc ggtacgcgta ttaattgcgt
     4021 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4201 tacggttatc cacagaatca ggggataacg caggaaag
//
