CaV1.2 vector (V000220#)

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

CaV1.211709 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500CAP binding sitelac promoterlac operatorM13 revSV40 poly(A) signalBSDEM7 promoterSV40 promoterpBABE 3'f1 oribGH poly(A) signal6xHisV5 tagRBST7CMV promoterCMV enhancerpRS-markerAmpR promoterAmpRoriL4440

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

Copy Sequence

Download GeneBank File(.gb)

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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