Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002459 | pCL-Eco | 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
- Vector Name:
- pCL-Eco
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12341 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMVd1
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- LXSN primer
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pCL-Eco 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.
pCL-Eco vector Sequence
LOCUS Exported 12341 bp ds-DNA circular SYN 20-DEC-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCL-Eco
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12341)
AUTHORS Naviaux RK, Costanzi E, Haas M, Verma IM
TITLE The pCL vector system: rapid production of helper-free, high-titer,
recombinant retroviruses.
JOURNAL J Virol. 1996 Aug . 70(8):5701-5.
PUBMED 8764092
REFERENCE 2 (bases 1 to 12341)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Virol.
1996 Aug . 70(8):5701-5."
FEATURES Location/Qualifiers
source 1..12341
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 133..152
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 386..403
/label=L4440
/note="L4440 vector, forward primer"
rep_origin 468..602
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(521..540)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 1060..1194
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1084..1103)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1138..1157
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1466..1486)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
intron 1616..1681
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS complement(4331..9547)
/codon_start=1
/gene="gag-pol"
/product="Moloney murine leukemia virus Gag-Pol
polyprotein"
/label=gag-pol
/note="contains a read-through stop codon"
/translation="MGQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPT
FNVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPK
PPPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGAKPKPQVLSDSGGPLIDLLTEDPP
PYRDPRPPPSDRDGNGGEATPAGEAPDPSPMASRLRGRREPPVADSTTSQAFPLRAGGN
GQLQYWPFSSSDLYNWKNNNPSFSEDPGKLTALIESVLITHQPTWDDCQQLLGTLLTGE
EKQRVLLEARKAVRGDDGRPTQLPNEVDAAFPLERPDWDYTTQAGRNHLVHYRQLLLAG
LQNAGRSPTNLAKVKGITQGPNESPSAFLERLKEAYRRYTPYDPEDPGQETNVSMSFIW
QSAPDIGRKLERLEDLKNKTLGDLVREAEKIFNKRETPEEREERIRRETEEKEERRRTE
DEQKEKERDRRRHREMSKLLATVVSGQKQDRQGGERRRSQLDRDQCAYCKEKGHWAKDC
PKKPRGPRGPRPQTSLLTLDD*GGQGQEPPPEPRITLKVGGQPVTFLVDTGAQHSVLTQ
NPGPLSDKSAWVQGATGGKRYRWTTDRKVHLATGKVTHSFLHVPDCPYPLLGRDLLTKL
KAQIHFEGSGAQVMGPMGQPLQVLTLNIEDEYRLHETSKEPDVSLGSTWLSDFPQAWAE
TGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQSPW
NTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVLDLKD
AFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFDEALHRDLADFRI
QHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQVKYLGY
LLKEGQRWLTEARKETVMGQPTPKTPRQLREFLGTAGFCRLWIPGFAEMAAPLYPLTKT
GTLFNWGPDQQKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVLTQKLGPWR
RPVAYLSKKLDPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHAVEALVKQPP
DRWLSNARMTHYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLDILAEAHGTRP
DLTDQPLPDADHTWYTDGSSLLQEGQRKAGAAVTTETEVIWAKALPAGTSAQRAELIAL
TQALKMAEGKKLNVYTDSRYAFATAHIHGEIYRRRGLLTSEGKEIKNKDEILALLKALF
LPKRLSIIHCPGHQKGHSAEARGNRMADQAARKAAITETPDTSTLLIENSSPYTSEHFH
YTVTDIKDLTKLGAIYDKTKKYWVYQGKPVMPDQFTFELLDFLHQLTHLSFSKMKALLE
RSHSPYYMLNRDRTLKNITETCKACAQVNASKSAVKQGTRVRGHRPGTHWEIDFTEIKP
GLYGYKYLLVFIDTFSGWIEAFPTKKETAKVVTKKLLEEIFPRFGMPQVLGTDNGPAFV
SKVSQTVADLLGIDWKLHCAYRPQSSGQVERMNRTIKETLTKLTLATGSRDWVLLLPLA
LYRARNTPGPHGLTPYEILYGAPPPLVNFPDPDMTRVTNSPSLQAHLQALYLVQHEVWR
PLAAAYQEQLDRPVVPHPYRVGDTVWVRRHQTKNLEPRWKGPYTVLLTTPTALKVDGIA
AWIHAAHVKAADPGGGPSSRLTWRVQRSQNPLKIRLTREAP"
misc_feature 4394..4768
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind complement(4440..4456)
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
primer_bind complement(9144..9160)
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(9176..9198)
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
promoter complement(9856..10059)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(9885..9905)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 10060..10439
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(10584..10603)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(10763..10781)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 10849..10953
/gene="bla"
/label=AmpR promoter
CDS 10954..11814
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(11172..11191)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin join(11985..12341,1..232)
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"