LOCUS       sequence-209229-        2916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knock down of hTERT transcript.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2916)
  AUTHORS   Kreilmeier T, Mejri D, Hauck M, Kleiter M, Holzmann K
  TITLE     Telomere Transcripts Target Telomerase in Human Cancer Cells.
  JOURNAL   Genes (Basel). 2016 Aug 16;7(8). pii: genes7080046. doi: 
            10.3390/genes7080046.
  PUBMED    27537914
REFERENCE   2  (bases 1 to 2916)
  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: "Genes 
            (Basel)."; date: "2016-08-16"; volume: "7(8). pii: genes7080046. 
            doi"; pages: " 10.3390/genes7080046"
COMMENT     Sequence Label: pENTR U6-hTERTsh
FEATURES             Location/Qualifiers
     source          1..2916
                     /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"
     promoter        720..968
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     720..740
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     891..910
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(1041..1140)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1157..1176)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1158..1176)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1181..1197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1181..1197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1299..1318)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1310..2119
                     /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(1377..1396)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2266..2854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2755..2774
                     /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 agcaggcttt aaaggaacca attcagtcga ctggatctgt cgacaaggtc gggcaggaag
      721 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      781 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      841 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      901 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
      961 gaaacaccgt ttcatcagca agtttggatt caagagatcc aaacttgctg atgaaatttt
     1021 tgaattctcg agatatctag acccagcttt cttgtacaaa gttggcatta taagaaagca
     1081 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1141 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     1201 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     1261 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1321 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1381 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     1441 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1501 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1561 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     1621 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1681 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1741 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1801 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     1861 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     1921 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     1981 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2041 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2101 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     2161 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     2221 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2281 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2341 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2401 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2461 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2521 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2581 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2641 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2701 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2761 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2821 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2881 gttcctggcc ttttgctggc cttttgctca catgtt
//
