pGFPuv

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  • Model: PVT10618
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pGFPuv


Catalog No. PVT10618
Packing 2ug

 

pGFPuv Information

Function Fluorescence E.coli Plasmid
Resistance Amp
Screen GFPuv
Strain DH5alpha
Culture temperature 37degrees centigrade
Replicon PUC
Copy high
Promoter Lac
Induction IPTG
Forward primer
Reverse primer

 

pGFPuv Description

PGFPuv is a green fluorescent expression plasmid of E. coli. pGFPuv carries the“cycle 3”variant of GFP described by Crameri et al. (1). This gene was cloned between the two MCSs of the pUC19 derivative pPD16.43 (2). The GFPuv gene canbe easily excised from pGFPuv. Alternatively, the GFPuv coding sequence can be amplified by PCR. The GFPuv gene was inserted in frame with the lacZ initiation codon from pUC19 so that a β-galactosidase-GFPuv fusionprotein is expressed from the lac promoter in E. coli . Note, however, that if you excise the GFPuv coding sequence using a restriction site in the 5' MCS, the resulting fragment will encode the native (i.e.,non-fusion) GFPuv protein. The pUC backbone of pGFPuv provides a high copy number origin of replication and ampicillin resistance gene for propagation in E. coli.

 

pGFPuv Multiple cloning site

pGFPuv-vector

 

pGFPuv Sequence

LOCUS       Exported                3337 bp ds-DNA     circular SYN 17-OCT-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..3337

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        143..173

                     /label=lac promoter

                     /note="promoter for the E. coli lac operon"

     protein_bind    181..197

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     primer_bind     205..221

                     /label=M13 rev

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     CDS             289..1005

                     /codon_start=1

                     /product="GFP variant optimized for excitation by UV light"

                     /label=GFPuv

                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK

                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG

                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA

                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE

                     FVTAAGITHGMDELYK"

     promoter        1432..1536

                     /gene="bla"

                     /label=AmpR promoter

     CDS             1537..2397

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      2568..3156

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 

                     replication"

ORIGIN

        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc

       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc

      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa

      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg

      241 catgcctgca ggtcgactct agaggatccc cgggtaccgg tagaaaaaat gagtaaagga

      301 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg

      361 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt

      421 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc

      481 tcttatggtg ttcaatgctt ttcccgttat ccggatcata tgaaacggca tgactttttc

      541 aagagtgcca tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg

      601 aactacaaga cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag

      661 ttaaaaggta ttgattttaa agaagatgga aacattctcg gacacaaact cgagtacaac

      721 tataactcac acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac

      781 ttcaaaattc gccacaacat tgaagatgga tccgttcaac tagcagacca ttatcaacaa

      841 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa

      901 tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta

      961 actgctgctg ggattacaca tggcatggat gagctctaca aataatgaat tccaactgag

     1021 cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggcct actagtcggc

     1081 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc

     1141 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc

     1201 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc

     1261 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa

     1321 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc

     1381 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc

     1441 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc

     1501 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc

     1561 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg

     1621 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat

     1681 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc

     1741 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa

     1801 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa

     1861 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt

     1921 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct

     1981 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat

     2041 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg

     2101 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg

     2161 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt

     2221 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg

     2281 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg

     2341 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg

     2401 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa

     2461 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt

     2521 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt

     2581 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt

     2641 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag

     2701 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta

     2761 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat

     2821 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg

     2881 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg

     2941 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac

     3001 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga

     3061 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt

     3121 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta

     3181 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat

     3241 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg

     3301 accgagcgca gcgagtcagt gagcgaggaa gcggaag

//

 

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