pAdEasy-1 vector (V012970) Gene synthesis in pAdEasy-1 backbone

Price Information

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V012970 pAdEasy-1 In stock, instant shipping

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

Basic Vector Information

pAdEasy-1 is an adenoviral vector plasmid derived from human adenovirus serotype 5 (Ad5), with the E1 and E3 genes deleted to accommodate foreign DNA inserts of up to 7.5 kb. It has a plasmid size of 33,477 bp and carries an ampicillin resistance gene for prokaryotic selection.

Vector Name:
pAdEasy-1
Antibiotic Resistance:
Ampicillin
Length:
33477 bp
Type:
Packaging assistance
Replication origin:
ori
Host:
Mammalian cells, Adenovirus
Source/Author:
Tong-Chuan He
Copy Number:
Low
Growth Strain(s):
DH10B
Growth Temperature:
30℃

pAdEasy-1 vector Map

pAdEasy-133477 bp1600320048006400800096001120012800144001600017600192002080022400240002560027200288003040032000POLL1Pre-hexon-linking protein IIIa from Human adenovirus C serotype 5. Accession#: P12537L2L2L2Pre-core protein X from Human adenovirus C serotype 5. Accession#: Q2KS10L3L3L3DBPL4L4Fiber protein from Human adenovirus C serotype 5. Accession#: P11818Early 4 ORF6 protein from Human adenovirus C serotype 2. Accession#: P03239Early 4 ORF3 protein from Human adenovirus C serotype 2. Accession#: P03241ITRropbomoriAmpRAmpR promoterIX

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. doi: 10.1073/pnas.95.5.2509. PMID: 9482916; PMCID: PMC19394.

pAdEasy-1 vector Sequence

LOCUS       Exported               33477 bp DNA     circular UNA 18-JUL-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 33477)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..33477
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     source          join(27380..33477,1..27379)
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     source          27382..28091
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     source          27382..28091
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     source          27382..28091
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     source          27382..28091
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    1584..1687
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     CDS             complement(3125..5128)
                     /gene="PTP"
                     /label=PTP
                     /note="Preterminal protein from Human adenovirus C serotype
                     5. Accession#: P04499"
     CDS             5587..6831
                     /gene="L1"
                     /label=L1
                     /note="Packaging protein 3 from Human adenovirus C serotype
                     5. Accession#: P04496"
     CDS             6855..8609
                     /note="Pre-hexon-linking protein IIIa from Human adenovirus
                     C serotype 5. Accession#: P12537"
     CDS             8693..10405
                     /gene="L2"
                     /label=L2
                     /note="Penton protein from Human adenovirus C serotype 5. 
                     Accession#: P12538"
     CDS             10415..11008
                     /gene="L2"
                     /label=L2
                     /note="Pre-histone-like nucleoprotein from Human adenovirus
                     C serotype 2. Accession#: P68950"
     CDS             11081..12184
                     /gene="L2"
                     /label=L2
                     /note="Core-capsid bridging protein from Human adenovirus C
                     serotype 5. Accession#: P24938"
     CDS             12215..12454
                     /note="Pre-core protein X from Human adenovirus C serotype
                     5. Accession#: Q2KS10"
     CDS             12540..13289
                     /gene="L3"
                     /label=L3
                     /note="Pre-protein VI from Human adenovirus C serotype 5. 
                     Accession#: P24937"
     CDS             13378..16233
                     /gene="L3"
                     /label=L3
                     /note="Hexon protein from Human adenovirus C serotype 5. 
                     Accession#: P04133"
     CDS             16269..16880
                     /gene="L3"
                     /label=L3
                     /note="Protease from Human adenovirus C serotype 5.
                     Accession#: P03253"
     CDS             complement(16982..18568)
                     /codon_start=1
                     /gene="DBP"
                     /label=DBP
                     /note="DNA-binding protein from Human adenovirus C serotype
                     5. Accession#: P03265"
                     /translation="MASREEEQRETTPERGRGAARRPPTMEDVSSPSPSPPPPRAPPKK
                     RMRRRIESEDEEDSSQDALVPRTPSPRPSTSAADLAIAPKKKKKRPSPKPERPPSPEVI
                     VDSEEEREDVALQMVGFSNPPVLIKHGKGGKRTVRRLNEDDPVARGMRTQEEEEEPSEA
                     ESEITVMNPLSVPIVSAWEKGMEAARALMDKYHVDNDLKANFKLLPDQVEALAAVCKTW
                     LNEEHRGLQLTFTSNKTFVTMMGRFLQAYLQSFAEVTYKHHEPTGCALWLHRCAEIEGE
                     LKCLHGSIMINKEHVIEMDVTSENGQRALKEQSSKAKIVKNRWGRNVVQISNTDARCCV
                     HDAACPANQFSGKSCGMFFSEGAKAQVAFKQIKAFMQALYPNAQTGHGHLLMPLRCECN
                     SKPGHAPFLGRQLPKLTPFALRNAEDLDADLISDKSVLASVHHPALIVFQCCNPVYRNS
                     RAQGGGPNCDFKISAPDLLNALVMVRSLWSENFTELPRMVVPEFKWSTKHQYRNVSLPV
                     AHSDARQNPFDF"
     CDS             18597..21017
                     /gene="L4"
                     /label=L4
                     /note="Shutoff protein from Human adenovirus C serotype 5. 
                     Accession#: P24933"
     CDS             21704..22384
                     /gene="L4"
                     /label=L4
                     /note="Pre-hexon-linking protein VIII from Human adenovirus
                     C serotype 5. Accession#: P24936"
     CDS             22939..24681
                     /note="Fiber protein from Human adenovirus C serotype 5. 
                     Accession#: P11818"
     CDS             complement(25093..25974)
                     /note="Early 4 ORF6 protein from Human adenovirus C
                     serotype 2. Accession#: P03239"
     CDS             complement(26253..26600)
                     /note="Early 4 ORF3 protein from Human adenovirus C
                     serotype 2. Accession#: P03241"
     repeat_region   27733..27835
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     CDS             29126..29317
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    29419..29559
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(29745..30333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(30504..31364)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(31365..31469)
                     /label=AmpR promoter
     CDS             31625..32044
                     /gene="IX"
                     /label=IX
                     /note="Hexon-interlacing protein from Human adenovirus C 
                     serotype 5. Accession#: P03281"
     CDS             complement(32110..33444)
                     /gene="IVa2"
                     /label=IVa2
                     /note="Packaging protein 1 from Human adenovirus C serotype
                     5. Accession#: P03271"
     CDS             complement(join(33216..33477,1..3323))
                     /gene="POL"
                     /label=POL
                     /note="DNA polymerase from Human adenovirus C serotype 2. 
                     Accession#: P03261"