pAC5.1b-EGFP
Search name
pAC5.1b-EGFP,Plasmid pAC5.1b-EGFP,pAC5.1b-EGFP vector
pAC5.1b-EGFP Information
Promoter: Ac5 promoter
Replicon: pUC ori
Terminator: SV40 poly (A) signal
Plasmid classification: insect series plasmids, insect free plasmids, insect intracellular plasmids.
Plasmid size: 6072bp
Plasmid Tags: N-EGFP, C-V5, C-6 x His
Prokaryotic resistance: ampicillin Ampicillin
Screening markers: no
Cloned strains of Escherichia coli, DH5 A and other Escherichia coli
Culture conditions: 37 centigrade, aerobic, LB
Expression host: Drosophila cell line S2
Induction mode: constituent expression without induction
5'sequencing primers: Ac5-F:ACACAAAGCCGCTCCATCAG
3'sequencing primers: BGH-R:TAGAAGGCACAGTCGAGG
Note: non virus
Use:Insect cell plasmid
pAC5.1b-EGFP Description
PAC5.1b-EGFP plasmid is an expression vector of an insect cell group. Ac5 promoter drives EGFP tags, target genes, Myc tags and 6 x HIS tags together to express them together. It can be co transfected with pCoHygro or pCoBlast and establish a stable expression cell line.
pAC5.1b-EGFP is a 6kb expression vector designed for use with the Drosophila Constitutive Expression System available from Invitrogen. Upon transfection, the vector allows transient expression of your protein of interest in Drosophila cells. When cotransfected with the selection vector, pCoHygro or pCoBlast, pAC5.1b-EGFP allows selection of stable cell lines exhibiting constitutive expression of the protein of interest.
• The Drosophila actin 5C (Ac5) promoter for high-level, constitutive expression of the gene of interest in S2 cells (Chung and Keller, 1990).
• Multiple cloning site to facilitate cloning the gene of interest.
• C-terminal peptide containing the V5 epitope and polyhistidine (6xHis) tag for detection and purification of your protein of interest (if desired).
• Three reading frames to facilitate in-frame cloning with the C-terminal peptide.
• N-terminal EGFP tag for better detection.
• Ampicillin resistance gene for selection of transformants in E. coli.

pAC5.1b-EGFP Sequence
LOCUS Exported 6072 bp ds-DNA circular SYN 23-MAY-2016
DEFINITION synthetic circular DNA
KEYWORDS pAC5.1b-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6072)
TITLE Direct Submission
JOURNAL Exported Monday, May 23, 2016
FEATURES Location/Qualifiers
source 1..6072
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 56..2568
/note="Ac5 promoter"
/note="Drosophila melanogaster actin 5C promoter"
CDS 2617..3339
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYKF"
CDS 3357..3398
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
CDS 3408..3425
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 3498..3632
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(4254..4842)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5013..5873)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5874..5978)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
2101 ccactataaa cacatgtaca tatgtatgtt tt