LOCUS       sequence-80283-e        3582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre/lox mammalian expression and gateway entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3582)
  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 3582)
  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.ON
FEATURES             Location/Qualifiers
     source          1..3582
                     /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"
     protein_bind    complement(1323..1356)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1453..1486)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1552..1569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1558..1782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1811..1910)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2022..2041)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2033..2842
                     /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(2100..2119)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2932..3520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3421..3440
                     /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 tggtttagtg aaccgtcaga tccgctagcg
     1321 ctataacttc gtatagcata cattatacga agttatagct cctaggaaat ttaaataagg
     1381 cgcgccaagt taacgggcgg ccgcatggat ccgccctcga gccgtttaaa caagaattcg
     1441 tcgagggacc taataacttc gtatagcata cattatacga agttatacat gtttaagggt
     1501 tccggttcca ctaggtacaa ttcgatagct taagtttaaa ccgctgatca gcctcgactg
     1561 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1621 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1681 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1741 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1801 ccagatctag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1861 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1921 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1981 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2041 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2101 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2161 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2221 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2281 gcattttatc cgtactcctg gtgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2341 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2401 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2461 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2521 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2581 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2641 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2701 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2761 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2821 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacatt attcagattg
     2881 ggccccgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3061 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3241 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3301 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3361 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3421 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3481 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3541 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
