LOCUS       sequence-8635-ex        2619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for EGFPshRNA#1 under H1/TO promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2619)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 2619)
  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: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pENTR/pTER shEGFP #1 (628-2)
FEATURES             Location/Qualifiers
     source          1..2619
                     /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"
     primer_bind     634..653
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    699..717
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(848..947)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1059..1078)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1070..1879
                     /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(1137..1156)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1969..2557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2458..2477
                     /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 ttcagtcgac
      481 tggatccggt accgaattcg aacgctgacg tcatcaaccc gctccaagga atcgcggccc
      541 agtgtcacta ggcgggaaca cccagcgcgc gtgcgccctg gcaggaagat ggctgtgagg
      601 gacaggggag tggcgccctg caatatttgc atgtcgctat gtgttctggg aaatcaccat
      661 aaacgtgaaa tgtctttgga tttgggaatc ttataagttc cctatcagtg atagagatcc
      721 cgaagcagca cgacttcttc ttcaagagag aagaagtcgt gctgcttctt tttggaaaag
      781 cttggtaccg agctcggatc cactagtcca gtgtggtgga attcgcggcc gcactcgaga
      841 tatctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
      901 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca
      961 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     1021 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1081 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1141 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     1201 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1261 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1321 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     1381 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1441 gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1501 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1561 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     1621 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     1681 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     1741 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     1801 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     1861 gctcgatgag tttttctaat cagaattggt taattggttg taacattatt cagattgggc
     1921 cccgttccac tgagcgtcag acccggtaga aaagatcaaa ggatcttctt gagatccttt
     1981 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2041 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2101 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2161 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2221 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2281 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2341 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2401 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2461 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2521 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2581 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
