pG5Luc
Search name
pG5Luc,Plasmid pG5Luc,pG5Luc vector
pG5Luc Plasmid information
Replicon: pUC ori, F1 ori
Terminator: SV40 poly (A) signal
Plasmid classification: mammalian cells, mammalian cells, two hybrid vectors
Plasmid size: 4955bp
Prokaryotic resistance: Amp
Clone strain: DH5 alpha
Culture conditions: 37 LB, aerobic
Expression host: mammalian cells
Induction method: no induction, transient expression
Primers for 5'sequencing: Sv40-polyA-R:GAAATTTGTGATGCTATTGC
Primers for 3'sequencing: primers were designed according to the sequence
pG5Luc Plasmid Description
The pG5luc Vector contains five GAL4 binding sites upstream of a minimal TATA box, which in turn is upstream of the firefly luciferase gene. The pGAL4 and pVP16 fusion constructs are transfected along with the pG5luc Vector into mammalian cells. Two to three days after transfection, the cells are lysed, and the amount of Renilla luciferase and firefly luciferase are quantitated using the Dual-Luciferase Reporter Assay System. Interaction between the two test proteins, expressed as GAL4 and VP16 fusion constructs, results in an increase in firefly luciferase expression as compared to the negative controls.
pG5Luc Plasmid Sequence
LOCUS Exported 4955 bp ds-DNA circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4955)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..4955
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 225..1877
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/note="luciferase"
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
polyA_signal 1918..2039
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2458..3046)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3217..4077)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4078..4182)
/gene="bla"
/note="AmpR promoter"
rep_origin 4209..4664
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal 4795..4843
/note="synthetic polyadenylation signal"
misc_feature 4857..4948
/note="pause site"
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
ORIGIN
1 ggtaccgagt ttctagacgg agtactgtcc tccgagcgga gtactgtcct ccgactcgag
61 cggagtactg tcctccgatc ggagtactgt cctccgcgaa ttccggagta ctgtcctccg
121 aagacgctag cggggggcta taaaaggggg tgggggcgtt cgtcctcact ctagatctgc
181 gatctaagta agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa
241 acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac
301 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac
361 atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag
421 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc
481 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga
541 acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg
601 tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca
661 tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca
721 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct
781 tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta
841 aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta
901 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg
961 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt
1021 atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc
1081 tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt
1141 tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag
1201 cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta
1261 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg
1321 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc
1381 aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg
1441 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact
1501 gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag
1561 gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg
1621 acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg
1681 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag
1741 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc
1801 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg
1861 gaaagatcgc cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa
1921 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
1981 gtgaaattt <