pAdEasy-2 vector (V005925)

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V005925 pAdEasy-2 In stock, instant shipping

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

Basic Vector Information

Adenoviral vector for use in AdEasy System, for use with large inserts.

Vector Name:
pAdEasy-2
Antibiotic Resistance:
Ampicillin
Length:
30808 bp
Type:
Adenoviral vectors
Replication origin:
ori
Copy Number:
Low copy number
Growth Strain(s):
DH10B
Growth Temperature:
37℃

pAdEasy-2 vector Vector Map

pAdEasy-230808 bp1500300045006000750090001050012000135001500016500180001950021000225002400025500270002850030000pLannotateYPB1_ECOLXropbomoriAmpRAmpR promoterHexon-interlacing proteinPackaging protein 1Preterminal proteinVAVAPackaging protein 3Pre-hexon-linking protein IIIaPenton proteinpLannotateCore-capsid bridging proteinPre-core protein XPre-protein VIHexon proteinProteaseDNA-binding proteinShutoff proteinpLannotatePre-hexon-linking protein VIIIpLannotateFiber proteinITR

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

  • He TC, Zhou S, da Costa LT, Yu J, Kinzler KW, Vogelstein B. A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2509-14.

pAdEasy-2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V005925                30808 bp    DNA     circular UNA 27-FEB-2024
DEFINITION  Exported.
ACCESSION   V005925
VERSION     V005925
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
            .
REFERENCE   1  (bases 1 to 30808)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Annotated with pLannotate v1.2.0
FEATURES             Location/Qualifiers
     source          1..30808
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             27..651
                     /codon_start=1
                     /label="TcR (fragment)"
                     /note="pLannotate"
                     /note="/fragment=True"
                     /note="/identity=99.8"
                     /note="/match_length=52.5"
                     /note="/other=CDS"
                     /translation="STDALESLQPSQLLPVGAGHDYRRRTYDCLLYHATRRTGAGSALG
                     HFRRGPLSLERDDDRPVACGIRNLARPRSSLRHWSRHQTFRREAGHYRRHGGRRAGLRL
                     AGVRDARLDGLPHYDSSRFRRHRDARVAGHAVQAGR*RPSGTASRIARGSYQPNFDHWT
                     ADRHGDLCRLGEHMERVGMDCRRRPIPCLPPRVASRCMEPGHLDL"
     CDS             complement(893..1288)
                     /codon_start=1
                     /label="YPB1_ECOLX"
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MPPCKGEFLFMGVMIPMKRERMLTIRVTDDEHARLLERCEGKQLA
                     VWMRRDQRKITQGQCQRFVNTDVGVPQGSQQHPAMQIRNIMVQGADFRVSRLYETRKPK
                     TIHVVAQVADVLQQQSLHVRSRIGDSFC"
     CDS             1292..1480
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    1585..1725
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1911..2499)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2673..3530)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3531..3635)
                     /label="AmpR promoter"
     CDS             3770..4189
                     /gene="IX"
                     /label="Hexon-interlacing protein"
                     /note="Hexon-interlacing protein from Human adenovirus C
                     serotype 5. Accession#: P03281"
     CDS             complement(4255..5589)
                     /gene="IVa2"
                     /label="Packaging protein 1"
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     CDS             complement(5361..8942)
                     /gene="POL"
                     /label="DNA polymerase"
                     /note="DNA polymerase from Human adenovirus C serotype 2.
                     Accession#: P03261"
     CDS             complement(8747..10750)
                     /gene="PTP"
                     /label="Preterminal protein"
                     /note="Preterminal protein from Human adenovirus C serotype
                     5. Accession#: P04499"
     ncRNA           10779..10935
                     /label="VA"
                     /note="pLannotate"
                     /note="/database=Rfam"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=98.7"
                     /note="/other=ncRNA"
     ncRNA           11034..11191
                     /label="VA"
                     /note="pLannotate"
                     /note="/database=Rfam"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=99.4"
                     /note="/other=ncRNA"
     CDS             11209..12453
                     /gene="L1"
                     /label="Packaging protein 3"
                     /note="Packaging protein 3 from Human adenovirus C serotype
                     5. Accession#: P04496"
     CDS             12477..14231
                     /note="Pre-hexon-linking protein IIIa from Human adenovirus
                     C serotype 5. Accession#: P12537"
                     /label="Pre-hexon-linking protein IIIa"
     CDS             14315..16027
                     /gene="L2"
                     /label="Penton protein"
                     /note="Penton protein from Human adenovirus C serotype 5.
                     Accession#: P12538"
     CDS             16037..16336
                     /codon_start=1
                     /label="L2 (fragment)"
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=100.0"
                     /note="/match_length=50.5"
                     /note="/other=CDS"
                     /translation="MSILISPSNNTGWGLRFPSKMFGGAKKRSDQHPVRVRGHYRAPWG
                     AHKRGRTGRTTVDDAIDAVVEEARNYTPTPPPVSTVDAAIQTVVRGARRYAKMKR"
     CDS             16702..17805
                     /gene="L2"
                     /label="Core-capsid bridging protein"
                     /note="Core-capsid bridging protein from Human adenovirus C
                     serotype 5. Accession#: P24938"
     CDS             17836..18075
                     /note="Pre-core protein X from Human adenovirus C serotype
                     5. Accession#: Q2KS10"
                     /label="Pre-core protein X"
     CDS             18161..18910
                     /gene="L3"
                     /label="Pre-protein VI"
                     /note="Pre-protein VI from Human adenovirus C serotype 5.
                     Accession#: P24937"
     CDS             19000..21855
                     /gene="L3"
                     /label="Hexon protein"
                     /note="Hexon protein from Human adenovirus C serotype 5.
                     Accession#: P04133"
     CDS             21891..22502
                     /gene="L3"
                     /label="Protease"
                     /note="Protease from Human adenovirus C serotype 5.
                     Accession#: P03253"
     CDS             complement(22604..24190)
                     /gene="DBP"
                     /label="DNA-binding protein"
                     /note="DNA-binding protein from Human adenovirus C serotype
                     5. Accession#: P03265"
     CDS             24219..26639
                     /gene="L4"
                     /label="Shutoff protein"
                     /note="Shutoff protein from Human adenovirus C serotype 5.
                     Accession#: P24933"
     CDS             26870..27241
                     /codon_start=1
                     /label="SF33K_ADE02 (fragment)"
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=96.8"
                     /note="/match_length=54.4"
                     /note="/other=CDS"
                     /translation="AHTAPAAAAAAATAAATQKQRRPDSKTLTKPKKSTAAAAAGGGAL
                     RLAPNEPVSTRELRNRIFPTLYAIFQQSRGQEQELKIKNRSLRSLTRSCLYHKSEDQLR
                     RTLEDAEALFSKYCALTLKD"
     CDS             27332..28012
                     /gene="L4"
                     /label="Pre-hexon-linking protein VIII"
                     /note="Pre-hexon-linking protein VIII from Human adenovirus
                     C serotype 5. Accession#: P24936"
     CDS             28016..28294
                     /codon_start=1
                     /label="E312_ADE02 (fragment)"
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=True"
                     /note="/identity=86.0"
                     /note="/match_length=86.9"
                     /note="/other=CDS"
                     /translation="MLSGEAEQLRLKHLVHCRRHKCFARDSGEFCYFELPEDHIEGPAH
                     GVRLTAQGELARSLIREFTQRPLLVERDRGPCVLTVICNCPNLGLHQD"
     CDS             28545..30287
                     /note="Fiber protein from Human adenovirus C serotype 5.
                     Accession#: P11818"
                     /label="Fiber protein"
     repeat_region   30704..30806
                     /label="ITR"
                     /note="inverted terminal repeat of human adenovirus
                     serotype 5"