| 载体基本信息 |
| 载体名称 |
pcDNA3.1(+) ZKSCAN1 MCS-ciRS7 Exon |
载体抗性 |
Ampicillin |
| 载体长度 |
8150 bp |
载体类型 |
Mammalian Expression |
| 启动子 |
CMV |
克隆方法 |
Restriction Enzyme |
| 5引物 |
CGCAAATGGGCGGTAGGCGTG |
pcDNA3.1(+) ZKSCAN1 MCS-ciRS7 Exon 载体载体图谱
pcDNA3.1(+) ZKSCAN1 MCS-ciRS7 Exon 载体载体序列
LOCUS Exported 8150 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses the human ciRS7/CDR1as circular RNA in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1(+) ZKSCAN1 MCS-ciRS7 Exon
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8150)
AUTHORS Kramer MC, Liang D, Tatomer DC, Gold B, March ZM, Cherry S, Wilusz
JE
TITLE Combinatorial control of Drosophila circular RNA expression by
intronic repeats, hnRNPs, and SR proteins.
JOURNAL Genes Dev. 2015 Oct 8.
PUBMED 26450910
REFERENCE 2 (bases 1 to 8150)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genes Dev.
2015 Oct 8."
FEATURES Location/Qualifiers
source 1..8150
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3744..3761)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3750..3974
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4020..4448
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4107..4126)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4317..4338
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4457..4477)
/label=pBABE 3
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4462..4791
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4642..4777
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4704..4723
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4858..5652
/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(4912..4931)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5522..5541
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5826..5947
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5863..5882)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5917..5936
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5996..6012)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5996..6012)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6009..6031)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6020..6036
/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(6044..6074)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6089..6110
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6227..6244)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6398..6983)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6478..6497)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7154..8014)
/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 7777..7796
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8015..8119)
/gene="bla"
/label=AmpR promoter