Basic Vector Information
- Vector Name:
- pEPIFOS-5
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 7518 bp
- Type:
- Cloning vector
- Replication origin:
- ori2
- Source/Author:
- EPICENTRE Biotechnologies.
pEPIFOS-5 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pEPIFOS-5 vector Sequence
LOCUS 40924_17586 7518 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pEPIFOS-5, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7518)
AUTHORS EPICENTRE Biotechnologies.
TITLE Direct Submission
JOURNAL Submitted (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA
REFERENCE 2 (bases 1 to 7518)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7518)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(23-AUG-2007) 726 Post Road, Madison, WI 53713, USA"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7518
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 288..304
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 311..329
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(443..459)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(467..483)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(491..521)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(536..557)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(780..1436)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(1437..1539)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin 2466..2685
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 2776..3528
/codon_start=1
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
misc_feature 3534..3784
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 4110..5282
/codon_start=1
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
CDS 5285..6253
/codon_start=1
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
misc_feature 6329..6802
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature complement(7062..7460)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind complement(7478..7511)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
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