pInSRT-GFPV1 vector (V005422)

Basic Vector Information

Vector Name:
pInSRT-GFPV1
Antibiotic Resistance:
Ampicillin
Length:
12929 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Wasa J, Nishida Y, Shinomura T, Urakawa H, Arai E, Futamura N, Tsukushi S, Ishiguro N.

pInSRT-GFPV1 vector Map

pInSRT-GFPV112929 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600SP6 promoterFRTsignal sequence derived from human PG-M/versicanEGFPIRES2PuroRbGH poly(A) signalT7 promoterM13 fwdf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 rev

pInSRT-GFPV1 vector Sequence

LOCUS       40924_25561       12929 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pInSRT-GFPV1 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12929)
  AUTHORS   Wasa J, Nishida Y, Shinomura T, Urakawa H, Arai E, Futamura N, 
            Tsukushi S, Ishiguro N.
  TITLE     Versican Regulates Cell-Associated Matrix Formation and Cell 
            Behavior Differentially from Aggrecan in Swarm Rat Chondrosarcoma 
            Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12929)
  AUTHORS   Shinomura T.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUN-2010) Contact:Tamayuki Shinomura Tokyo Medical and
            Dental University, Tissue Regeneration; 1-5-45 Yushima, Bunkyo-Ku, 
            Tokyo 113-8549, Japan
REFERENCE   3  (bases 1 to 12929)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12929)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JUN-2010) Contact:Tamayuki Shinomura Tokyo Medical and Dental 
            University, Tissue Regeneration; 1-5-45 Yushima, Bunkyo-Ku, Tokyo 
            113-8549, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..32
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    50..97
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    177..236
                     /gene="GFPV1"
                     /label=signal sequence derived from human PG-M/versican
                     /note="signal sequence derived from human PG-M/versican"
     CDS             279..995
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    8297..8883
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             8884..9480
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    9570..9794
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(9828..9846)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9853..9869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(10010..10465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        10799..10903
                     /label=AmpR promoter
     CDS             10904..11761
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      11935..12523
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    12811..12832
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        12847..12877
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12885..12901
                     /label=lac operator
                     /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     12909..12925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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