Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012547 | pBeloBAC11 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
pBeloBAC11 is a single-copy E. coli plasmid vector designed for constructing Bacterial Artificial Chromosomes (BACs)
- Vector Name:
- pBeloBAC11
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 7507 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori2
- Source/Author:
- New England Biolabs
- Copy Number:
- Low copy number
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 30℃
pBeloBAC11 vector Vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Ávila-Pérez G, Park JG, Nogales A, Almazán F, Martínez-Sobrido L. Rescue of Recombinant Zika Virus from a Bacterial Artificial Chromosome cDNA Clone. J Vis Exp. 2019 Jun 24;(148).
pBeloBAC11 vector Sequence
LOCUS 40924_6182 7507 bp DNA circular SYN 01-JAN-1980
DEFINITION Single-copy E. coli plasmid vector designed for constructing
Bacterial Artificial Chromosomes (BACs).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7507)
AUTHORS New England Biolabs
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7507)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7507
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 289..305
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 312..330
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 333..389
/label=MCS
/note="pUC18/19 multiple cloning site"
promoter complement(396..414)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(432..448)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(456..472)
/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(480..510)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(525..546)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(769..1425)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(1426..1528)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin 2455..2674
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 2765..3517
/codon_start=1
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
misc_feature 3523..3773
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 4099..5271
/codon_start=1
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
CDS 5274..6242
/codon_start=1
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
misc_feature 6318..6791
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature complement(7051..7449)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind complement(7467..7500)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."