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pTransgene vector (V002558#)

Basic Vector Information

Basic Vector Information
Vector Name pTransgene Antibiotic Resistance Ampicillin
Length 5398 bp Type Cloning vector
Source Yang X, Pachikara B, Murray TJ, Liu D, Sonenshein GE, Sherr DH.

pTransgene vector Vector Map

BglII, NheI, XhoI and EcoRI restriction sites to isolate transgene construct chicken beta-globin core insulator PacI and AvrII restriction sites for changing promoter SacII and SpeI restriction sites for changing promoter chimeric intron:5'-donor site is from the 1st intronof the human beta-globin gene, the branch and 3'-acceptor site is from the intron that is between the leader and the body of an immunoglobulin gene heavy chain variable region multiple cloning site SV40 late polyadenylation signal chicken beta-globin core insulator SphI and SbfI restriction sites to isolate transgeneconstruct f1 ori AmpR promoter ampR ori

Plasmid Resuspension 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 ℃.

pTransgene vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5398 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTransgene, complete sequence.
ACCESSION   AF515846
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5398)
  AUTHORS   Yang X, Pachikara B, Murray TJ, Liu D, Sonenshein GE, Sherr DH.
  TITLE     pTransgene: A cloning vector with insulators for optimal cDNA 
            transgene expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5398)
  AUTHORS   Yang X, Sonenshein GE, Sherr DH.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAY-2002) Environmental Health, Boston University 
            School of Public Health and Medicine, 715 Albany Street, R-405, 
            Boston, MA 02118, USA
REFERENCE   3  (bases 1 to 5398)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5398
                     /organism="Cloning vector pTransgene"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:198113"
     misc_feature    1..56
                     /note="BglII, NheI, XhoI and EcoRI restriction sites to 
                     isolate transgene construct"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    62..635
                     /note="chicken beta-globin core insulator; shields 
                     transgene from chromosomal positional effects"
     misc_feature    647..707
                     /note="PacI and AvrII restriction sites for changing 
                     promoter"
     regulatory      668..1948
                     /regulatory_class="promoter"
                     /note="mouse mammary tumor virus 5' LTR"
     misc_feature    1949..1960
                     /note="SacII and SpeI restriction sites for changing 
                     promoter"
     intron          1961..2145
                     /note="chimeric intron:5'-donor site is from the 1st intron
                     of the human beta-globin gene, the branch and 3'-acceptor 
                     site is from the intron that is between the leader and the 
                     body of an immunoglobulin gene heavy chain variable region;
                     increases the level of transgene expression"
     intron          1986..2118
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    2146..2181
                     /note="multiple cloning site; contains MluI, XbaI, SalI, 
                     AccI, EagI and NotI restriction sites"
     regulatory      2182..2419
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal; enhance RNA 
                     stability and translation"
     polyA_signal    2197..2318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2424..2678
                     /note="chicken beta-globin core insulator; shields 
                     transgene from chromosomal positional effects"
     misc_feature    2702..2714
                     /note="SphI and SbfI restriction sites to isolate transgene
                     construct"
     rep_origin      2799..3254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3586..3690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3691..4551
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicillin resistance"
                     /protein_id="AAM70477.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3691..4551
                     /gene="ampR"
                     /label=ampR
     rep_origin      4722..5310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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