Mossing Lab - DNA sequences

Mossing lab DNA sequences.

Cloning Vectors

• pUC18.fasta Here is a link to the official pUC18 genbank entry at NCBI.
• pUC118.fasta Official pUC118 genbank entry at NCBI.
• pET21a PDF Map from Novagen
• pET33b PDF Map from Novagen

Cro Expression plasmids

• Here is a graphical genealogy Cro Plasmid Maps (PDF) of Cro expression plasmids from the Sauer Lab
• pUCroRS.fasta and Reverse Complement (pUCroRSrc.fasta) EcoRI - Tac promoter and synthetic Cro gene - (SmaI) of pAP120 inserted in pUC18. Interactive HTML from sequence extractor.
• Cro substitution mutant - tryptophan for phenylalanine at 58 Reverse Complement pUCroF58W_RSrc.fasta
• pUC118CroRX like pUCroRS but includes terminator from pAP120. This PDF file shows the difference.
• pUC119CroRX like pUC118CroRX but EcoR1 to hindIII segment is reversed. This PDF file shows the difference.
• pUCromDG.fasta like pUCroRs but contains a small insertion in the Cro gene. Mossing & Sauer Science (1990)
• Cro K56[DGEVK] F58W fragment (EcoRI) - Tac promoter and synthetic Cro monomer gene.
• pJF1a Cro in pET21a with C-terminal His 6 fusion (91 Amino Acids)
• pscCro8 single chain dimer, 8 residue linker in pET21a (reverse complement) Interactive HTML from sequence extractor.
• pscCro16 single chain dimer, 16 residue linker in pET21a (reverse complement).
• pET33bscCro8.fasta Reverse complement of pET33b sequence above with scCro 8 sequence cloned between NdeI and XhoI sites.

Reporter constructs

• pucOR_Cro lambda OR-Olac control region replaces the Tac promoter of puCroRS
• pUCOR_Cro_lac same as pucOR_Cro but with translation start of lac Zalpha reconstructed. Here is an annotated plain text sequence file of the important parts.
• pAH125 (PDF summary pAH125.pdf | genbank sequencepAH125.genbank | ncbi link) is a promoterless lacZ reporter plasmid, with kan^R, OriR and Att lambda for generation of single copy integrants
• pUA139 is similar to pMS201 (Map and sites pMS201.html | genbank sequence pMS201.genbank) except BamHI and XhoI sites upstream of GFPmut2 are reversed (Bam-Xho-GFP), kan^R and pSC101 origin
• pUAL306 (guesstimate PDF summary pUALuxBX.pdf | guesstimate fasta sequence pUALuxBX.fasta ) BamHI and XhoI sites upstream of has luxCDABE kan^R and pSC101 origin
• Hybrid P_R-lacO promoter oligonucleotides PR-lacO_oligos.txt
• New plasmid constructed by Yin Gu from pUA139 and Hybrid P_R-lacO promoter oligonucleotides pOR1GFP.fasta.
• Raw sequence data from 9/8/05.
• pOR1GFP + scCro DNA pOR1scCroGFP.fasta.
• pOR1GFP + scCro DNA reversed pOR1scCro_RGFP.fasta.
• pAH125 Pst-Sal-lacI(rc)-Sph-OPO-Xba...-EcoRI-BamHI-lacZ pAH125laciOPO.fasta. lacI (rev. comp.) + lambda PR/lac O promoter inserted between the PstI and XbaI sites of pAH125 Fixed 2016.03.07
• pAH125 Pst-Sal-lacI(rc)-Sph-OPOXba-scCro-XhoI-EcoRI-BamHI-lacZ pAH125lacI_OPO_scCro.fasta.

Last Updated: Tuesday, July 26, 2005

Online DNA sequence analysis resources

• VectorEditor from the Joint BioEnergy Institute (JBEI) restriction sites and open reading frames and draws maps.
• Nebcutter at New England Biolabs finds restriction sites and open reading frames and draws maps.
• Sequence Extractor generates an interactive annotated html file showing the locations of primers, restriction sites, and protein translations. Primers in SE format.
• PlasMapper automatically generates and annotates graphical plasmid maps in PNG, JPG, SVG or SVGZ format from DNA sequence.
• Primer3 picks primers for PCR
• Ape A plasmid editor - download for Mac, Linux or Windows

Links

• University of Mississippi
• Department of Chemistry
• Chemistry Faculty Page

References

• CRIM Plasmid/Host system. Haldimann and Wanner PDF
• Reference: Xiaoli Dong, Paul Stothard, Ian J. Forsythe, and David S. Wishart "PlasMapper: a web server for drawing and auto-annotating plasmid maps" Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W660-4.