Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | CaV1.2 | Antibiotic Resistance | Ampicillin |
| Length | 11709 bp | Type | Mammalian Expression |
| Cloning Method | Restriction Enzyme | 5' Primer | T7 |
| 3' Primer | BGH-rev | ||
CaV1.2 vector Vector Map
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 ℃.
CaV1.2 vector Sequence
LOCUS Exported 11709 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS CaV1.2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11709)
AUTHORS Helton TD, Xu W, Lipscombe D
TITLE Neuronal L-type calcium channels open quickly and are inhibited
slowly.
JOURNAL J Neurosci. 2005 Nov 2. 25(44):10247-51.
PUBMED 16267232
REFERENCE 2 (bases 1 to 11709)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Neurosci.
2005 Nov 2. 25(44):10247-51."
FEATURES Location/Qualifiers
source 1..11709
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 270..391
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(281..300)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 335..354
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(549..947)
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
promoter complement(966..1013)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(1061..1257)
/label=SV40 promoter
/note="SV40 early promoter"
promoter complement(1069..1279)
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 1075..1210
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1129..1148)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 1303..1323
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(1332..1760)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1442..1463)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1654..1673
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 1806..2030
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 2019..2036
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(2059..2076)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(2086..2127)
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
RBS 6340..6351
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
primer_bind complement(8891..8910)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8892..8910)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(8955..9158)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(8984..9004)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 9159..9538
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 9710..9729
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 9804..9908
/gene="bla"
/label=AmpR promoter
CDS 9909..10769
/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"
primer_bind complement(10127..10146)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10940..11528
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11429..11448
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11682..11699
/label=L4440
/note="L4440 vector, forward primer"
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