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pYSEAP vector (V001669)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001669 pYSEAP In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pYSEAP
Antibiotic Resistance:
Blasticidin
Length:
5298 bp
Type:
Mammalian Expression
Replication origin:
ori
Selection Marker:
Blasticidin
Promoter:
EF-1α
5' Primer:
NA
3' Primer:
NA

pYSEAP vector Map

Copy Sequence View Map
pYSEAP5298 bp6001200180024003000360042004800SV40 promoterEF-1-alpha promoterSEAPSV40 poly(A) signaloriBSDEM7 promoter

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.

pYSEAP vector Sequence

LOCUS       40924_48273        5298 bp DNA     circular SYN 27-JUL-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5298)
  AUTHORS   Pastrana DV, Buck CB, Pang YY, Thompson CD, Castle PE, FitzGerald 
            PC, Kruger Kjaer S, Lowy DR, Schiller JT
  TITLE     Reactivity of human sera in a sensitive, high-throughput 
            pseudovirus-based papillomavirus neutralization assay for HPV16 and 
            HPV18.
  JOURNAL   Virology. 2004 Apr 10;321(2):205-16.
  PUBMED    15051381
REFERENCE   2  (bases 1 to 5298)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5298)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Virology.";
            date: "2004-04-10"; volume: "321(2)"; pages: "205-16"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        681..1859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1940..3448
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="PTMLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKL
                     QPAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNV
                     DKHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVG
                     VVTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGG
                     RKYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSV
                     THLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHE
                     SRAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPG
                     KARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAV
                     FARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    complement(3506..3627)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4046..4634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4666..5061)
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(5080..5127)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     promoter        complement(join(5175..5298,1..206))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"