LOCUS       sequence-10198-e        2981 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2981)
  TITLE     shMDC1 plasmids
REFERENCE   2  (bases 1 to 2981)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pENTR/shp16 (w98-1)
FEATURES             Location/Qualifiers
     source          1..2981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    489..513
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        535..553
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     535..552
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        814..1062
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     814..834
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     985..1004
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(1148..1172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1210..1309)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1421..1440)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1432..2241
                     /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
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1499..1518)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2331..2919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2820..2839
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgag
      481 gttatcaaac aagtttgtac aaaaaagcag gctccgcggc cgcccccttc accgatttag
      541 gtgacactat agaatacaag cttggctgca ggtcgacgga tccccccgag tccaacaccc
      601 gtgggaatcc catgggcacc atggcccctc gctccaaaaa tgctttcgcg tcgcgcagac
      661 actgctcggt agtttcgggg atcagcgttt gagtaagagc ccgcgtctga accctccgcg
      721 ccgccccggc cccagtggaa agacgcgcag gcaaaacgca ccacgtgacg gagcgtgacc
      781 gcgcgccgag cgcgcgccaa ggtcgggcag gaagagggcc tatttcccat gattccttca
      841 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
      901 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
      961 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
     1021 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgaccgtaa ctattcggtg
     1081 cgttgggcag aagcttgtgc tcaacgcacc gaatagttgc ggtctttttt gaagggtggg
     1141 cgcgccgacc caactttctt gtacaaagtg gttcgataac gaattcgcgg ccgcactcga
     1201 gatatctaga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1261 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1321 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1381 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1441 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1501 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1561 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1621 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1681 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1741 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1801 gtgtccctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1861 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1921 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     1981 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2041 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2101 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2161 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2221 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg
     2281 gccccgttcc actgagcgtc agacccggta gaaaagatca aaggatcttc ttgagatcct
     2341 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2401 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2461 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2521 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2581 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2641 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2701 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2761 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2821 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2881 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2941 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
