pOKE104 vector (V004235)

MSDS

Basic Vector Information

Vector Name:: pOKE104

Antibiotic Resistance:: Ampicillin

Length:: 5165 bp

Type:: Transformation vector

Replication origin:: ori

Source/Author:: Low W, Jedd G.

Promoter:: SP6

pOKE104 vector Vector Map

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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 ℃.

pOKE104 vector Sequence

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LOCUS       40924_33747        5165 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pOKE104, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5165)
  AUTHORS   Low W, Jedd G.
  TITLE     An improved plasmid for transformation of Neurospora crassa using 
            the pan-2 gene as a selectable marker
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5165)
  AUTHORS   Low W, Jedd G.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-2008) Temasek Life Sciences Laboratory, 1 Research
            Link National University of Singapore 117604, Singapore
REFERENCE   3  (bases 1 to 5165)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5165)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-DEC-2008) Temasek Life Sciences Laboratory, 1 Research Link 
            National University of Singapore 117604, Singapore"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..61
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(68..86)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(104..120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(128..144)
                     /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)."
     promoter        complement(152..182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(197..218)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(506..1094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1268..2125)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2126..2230)
                     /label=AmpR promoter
     CDS             2889..2912
                     /codon_start=1
                     /label=8xHis
                     /note="8xHis affinity tag"
                     /translation="HHHHHHHH"
     CDS             join(2989..3255,3473..4135)
                     /codon_start=1
                     /gene="pan-2"
                     /product="3-methyl-2-oxobutanoate hydroxymethyltransferase"
                     /label=pan-2
                     /note="pantothenic acid-2"
                     /protein_id="ACM43517.1"
                     /translation="MGLPPTNPRKKVTLNTLQSMYKKGEKITMITAHDFPSAHVAEHAG
                     MDMILVGDSLAMVALGMEDTSEVLVEEMLLHCRSVARATSRAFTVGDLPMGSYEISPQQ
                     ALSTAIRFIKEGRVSAVKLEGGAEMAPTIRAITTAGIPVLAHIGLTPQRQNALGGFRVQ
                     GKSKESAKRLLRDALAVQEAGAFAVVVEAVPQEVAAFITGLLKIPTIGIGAGNGTSGQV
                     LVQADMTGNFPPGRFLPKFVKKYGDVWGEALRAIETYRDEVKGGSYPAEEHTYPMAKEE
                     VEAFVGDVRRLMNANASAKAEGEGEKDA"
     gene            2989..4135
                     /gene="pan-2"
                     /label=pan-2
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5126..5142
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5149..5165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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