LOCUS       sequence-129524-        4051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  intronically embedded eSIBR backbone in gateway middle entry vector 
            (pDONR221).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4051)
  AUTHORS   Fowler DK, Williams C, Gerritsen AT, Washbourne P
  TITLE     Improved knockdown from artificial microRNAs in an enhanced miR-155 
            backbone: a designer's guide to potent multi-target RNAi.
  JOURNAL   Nucleic Acids Res. 2015 Nov 17. pii: gkv1246.
  PUBMED    26582923
REFERENCE   2  (bases 1 to 4051)
  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: "Nucleic 
            Acids Res. 2015 Nov 17. pii: gkv1246."
COMMENT     Sequence Label: pME-eSIBR
FEATURES             Location/Qualifiers
     source          1..4051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     1262..1281
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(2176..2275)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2292..2311)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2293..2311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2316..2332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2316..2332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2434..2453)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2445..3254
                     /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     complement(2512..2531)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3401..3989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3890..3909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
      721 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
      781 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
      841 tggcggctgt tcccgagtct tgaatagaac cttcgctaat gcgggaaagc tcttattcgg
      901 gtgagatggg ctggggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
      961 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     1021 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     1081 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     1141 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     1201 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     1261 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     1321 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt
     1381 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacgaagc taaggcgcgc
     1441 ctgagaactt cagggtgagt ttggggaccc ttgattgttc tttctttttc gctattgtaa
     1501 aattcatgtt atatggaggg ggcaaagttt tcagggtgtt gtttagaatg ggaagatgtc
     1561 ccttgtatca ccatgcatgg accctcatga taattttgtt tctttcactt tctactctgt
     1621 tgacaaccat tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt
     1681 agcttgcatt tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagatc
     1741 ccatcgattc gaattcaaag tcgactggag gcttgctttg ggctgtatgc tgttgtcttc
     1801 aagatctgga agacacagga cacaaggccc tttatcagca ctcacatgga acaaatggcc
     1861 accgtgggag gatgacaact cgagcctcta gacaatcagg gtatattata ttgtacttca
     1921 gcacagtttt agagaacaat tgttataatt aaatgataag gtagaatatt tctgcatata
     1981 aattctggct ggcgtggaaa tattcttatt ggtagaaaca actacatcct ggtcatcatc
     2041 ctgcctttct ctttatggtt acaatgatat acactgtttg agatgaggat aaaatactct
     2101 gagtccaaac cgggcccctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     2161 ctgggcaacg tgctgaccca gctttcttgt acaaagttgg cattataaga aagcattgct
     2221 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     2281 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2341 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     2401 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     2461 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     2521 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     2581 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     2641 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     2701 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     2761 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     2821 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     2881 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     2941 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     3001 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3061 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3121 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3181 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3241 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     3301 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4021 tggccttttg ctggcctttt gctcacatgt t
//
