| 载体基本信息 | |||
| 载体名称 | pcDNA-K-GECO1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6598 bp | 载体类型 | Mammalian Expression |
| 拷贝数 | High Copy | ||
pcDNA-K-GECO1 载体载体图谱
pcDNA-K-GECO1 载体载体序列
LOCUS Exported 6598 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Mammalian cell expression plasmid for red fluorescent Ca2+ indicator
K-GECO1.
ACCESSION .
VERSION .
KEYWORDS pcDNA-K-GECO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6598)
AUTHORS Shen Y, Dana H, Abdelfattah AS, Patel R, Shea J, Molina RS, Rawal B,
Rancic V, Chang YF, Wu L, Chen Y, Qian Y, Wiens MD, Hambleton N,
Ballanyi K, Hughes TE, Drobizhev M, Kim DS, Koyama M, Schreiter ER,
Campbell RE
TITLE A genetically encoded Ca(2+) indicator based on circularly
permutated sea anemone red fluorescent protein eqFP578.
JOURNAL BMC Biol. 2018 Jan 16;16(1):9. doi: 10.1186/s12915-018-0480-0.
PUBMED 29338710
REFERENCE 2 (bases 1 to 6598)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC Biol.";
date: "2018-01-16"; volume: "16(1):9. doi"; pages: "
10.1186/s12915-018-0480-0"
FEATURES Location/Qualifiers
source 1..6598
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 110..489
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 490..693
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 644..664
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 738..757
/label=T7
/note="T7 promoter, forward primer"
promoter 738..756
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 781..790
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(2064..2081)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2070..2294
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2340..2768
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2427..2446)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2637..2658
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2777..2797)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2782..3111
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2962..3097
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3024..3043
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3178..3972
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3232..3251)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3842..3861
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4146..4267
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4183..4202)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4237..4256
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4316..4332)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4316..4332)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4329..4351)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4340..4356
/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)."
promoter complement(4364..4394)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4409..4430
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4547..4564)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4718..5306)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4798..4817)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5477..6337)
/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 6100..6119
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6338..6442)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6517..6536)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
我们不提供该载体，载体信息仅供查询和参考。