LOCUS       sequence-80284-e        3495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression and gateway entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3495)
  AUTHORS   Radulovich N, Leung L, Tsao MS
  TITLE     Modified gateway system for double shRNA expression and Cre/lox 
            based gene expression.
  JOURNAL   BMC Biotechnol. 2011 Mar 22;11:24.
  PUBMED    21418658
REFERENCE   2  (bases 1 to 3495)
  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: "BMC 
            Biotechnol."; date: "2011-03-22"; volume: "11"; pages: "24"
COMMENT     Sequence Label: pENTR.CMV
FEATURES             Location/Qualifiers
     source          1..3495
                     /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     complement(517..536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        708..1087
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1088..1291
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1242..1262
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1336..1355
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1336..1354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1465..1482)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1471..1695
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1724..1823)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1935..1954)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1946..2755
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2013..2032)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2845..3433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3334..3353
                     /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 gcaggctcca ccatgggaac caattcagtc
      481 gactggatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
      541 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      601 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      661 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      721 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      781 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      841 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      901 tcaatgggtg gactatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      961 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     1021 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     1081 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     1141 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     1201 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     1261 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     1321 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agcgtttaaa
     1381 cgggccctct agactcgaaa ttccaccaca ctggactagt ggatccgagc tcggtaccaa
     1441 gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1501 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1561 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1621 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1681 cggtgggctc tatggcttct gaggcggaaa gaaccagatc tagacccagc tttcttgtac
     1741 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1801 agtcaaaata aaatcattat ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct
     1861 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1921 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     1981 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2041 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2101 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2161 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctggtgatgc
     2221 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2281 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2341 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2401 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2461 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2521 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2581 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2641 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2701 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2761 attggttaat tggttgtaac attattcaga ttgggccccg ttccactgag cgtcagaccc
     2821 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2881 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2941 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3001 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3061 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3121 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3181 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3241 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3301 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3361 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3421 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3481 ttttgctcac atgtt
//
