PBDGTV vector (V009238)

Basic Vector Information

      • Vector Name:
      • PBDGTV
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7640 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Liu G, Wang X, Liu Y, Zhang M, Jia Y, Huang Y.

PBDGTV vector Vector Map

PBDGTV7640 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500AmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revT3 promoterCcdc107PGK promoter; phosphoglycerate kinase 1vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)bGH poly(A) signalDom3zT7 promoterM13 fwdf1 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 ℃.

PBDGTV vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7640 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector PBDGTV, complete sequence.
ACCESSION   KU179219
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7640)
  AUTHORS   Liu G, Wang X, Liu Y, Zhang M, Jia Y, Huang Y.
  TITLE     Arrayed mutant haploid stem cell libraries facilitate 
            phenotype-driven genetic screens
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7640)
  AUTHORS   Liu G, Wang X, Liu Y, Zhang M, Jia Y, Huang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-NOV-2015) Department of Medical Genetics, Institute of
            Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking
            Union Medical College, dong dan san tiao 5 hao, Beijing, Beijing 
            100005, China
REFERENCE   3  (bases 1 to 7640)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7640
                     /organism="Cloning vector PBDGTV"
                     /mol_type="other DNA"
                     /db_xref="taxon:1928191"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     mobile_element  complement(2248..2557)
                     /mobile_element_type="transposon:PB3'TR"
     gene            2569..3292
                     /gene="Ccdc107"
                     /label=Ccdc107
     exon            2616..2728
                     /gene="Ccdc107"
                     /note="penultimate Ccdc107 exon"
     exon            2804..2927
                     /gene="Ccdc107"
                     /note="final Ccdc107 exon"
     regulatory      3315..3827
                     /regulatory_class="promoter"
                     /note="PGK promoter; phosphoglycerate kinase 1"
     promoter        3320..3819
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      3896..3905
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3902..5503
                     /codon_start=1
                     /product="puro-deltaTK"
                     /label=puro-deltaTK
                     /note="puromycin resistant gene and deltaTK fusion protein"
                     /protein_id="APU57728.1"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAGSMPTLLRVYIDGPHGMGKTTT
                     TQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQHRLDQGEISAGDAAVVMTSA
                     QITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDRHPIAALLCYPAARYLMGSMT
                     PQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLL
                     ANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPN
                     GDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPT
                     ICDLARTFAREMGEAN"
     CDS             3902..4501
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             4508..5503
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=-Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    5531..5755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     gene            complement(6144..6561)
                     /gene="Dom3z"
                     /label=Dom3z
     exon            complement(6144..6387)
                     /gene="Dom3z"
                     /note="final Dom3z exon"
     exon            complement(6467..6561)
                     /gene="Dom3z"
                     /note="penultimate Dom3z exon"
     mobile_element  6686..6920
                     /mobile_element_type="transposon:PB5'TR"
     promoter        complement(7000..7018)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7025..7041)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7183..7638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"

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