| 载体基本信息 | |||
| 载体名称 | pOpt_Clover_Hyg | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6831 bp | 载体类型 | Cloning vector |
| 载体来源 | Lauersen KJ, Mussgnug JH, Kruse O. | ||
pOpt_Clover_Hyg 载体载体图谱
pOpt_Clover_Hyg 载体载体序列
LOCUS Exported 6831 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pOpt_Clover_Hyg, complete sequence.
ACCESSION KM061067
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6831)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Efficient targeted expression of nuclear transgenes in Chlamydomonas
reinhardtii with a novel, modular vector toolkit
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6831)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Direct Submission
JOURNAL Submitted (25-JUN-2014) Algae Biotechnology and Bioenergy,
University of Bielefeld Center for Biotechnology, Universitaetsstr
27, Bielefeld, NRW 33615, Germany
REFERENCE 3 (bases 1 to 6831)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Direct Submission
JOURNAL Submitted (18-MAY-2016) Algae Biotechnology and Bioenergy,
University of Bielefeld Center for Biotechnology, Universitaetsstr
27, Bielefeld, NRW 33615, Germany
REFERENCE 4 (bases 1 to 6831)
AUTHORS .
TITLE Direct Submission
COMMENT On May 18, 2016 this sequence version replaced KM061067.1.
##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6831
/organism="Cloning vector pOpt_Clover_Hyg"
/mol_type="other DNA"
/db_xref="taxon:1549205"
misc_feature 1..577
/label=pBluescript II KS(+)
/note="pBluescript II KS(+)"
primer_bind 489..505
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 515..533
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 578..844
/regulatory_class="promoter"
/note="Chlamydomonas reinhardtii heat shock protein 70A
(HSP70Ap) promoter"
regulatory 851..1074
/regulatory_class="promoter"
/note="Chlamydomonas reinhardtii ribulose-1,5-bisphosphate
carboxylase/oxygenase small subunit 2 promoter (RBCS2p)"
CDS join(1072..1079,1225..1658,1988..2319)
/codon_start=1
/product="clover green fluorescent protein"
/label=clover green fluorescent protein
/note="contains carboxy-terminal StrepII tag"
/protein_id="AIS68762.1"
/translation="MARMHMRSDVIEGRVSKGEELFTGVVPILVELDGDVNGHKFSVRG
EGEGDATNGKLTLKFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPE
GYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHN
VYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALS
KDPNEKRDHMVLLEFVTAAIEGRDIEFWSHPQFEK"
intron 1080..1224
/note="Chlamydomonas reinhardtii ribulose-1,5-bisphosphate
carboxylase/oxygenase small subunit 2 intron 1 (RBCS2i1)"
CDS 1247..1258
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
intron 1659..1987
/note="Chlamydomonas reinhardtii ribulose-1,5-bisphosphate
carboxylase/oxygenase small subunit 2 intron 2 (RBCS2i2)"
CDS 2269..2280
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 2293..2316
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
3'UTR 2320..2553
/note="Chlamydomonas reinhardtii ribulose-1,5-bisphosphate
carboxylase/oxygenase small subunit 2 3'UTR"
misc_feature 2554..2611
/label=multiple cloning site
/note="multiple cloning site"
regulatory 2612..2878
/regulatory_class="promoter"
/note="HSP70Ap"
regulatory 2885..3105
/regulatory_class="promoter"
/note="RBCS2p"
intron 3114..3258
/note="RBCS2i1"
CDS 3266..4264
/codon_start=1
/gene="aphVII"
/product="aminoglycoside 3'-phosphotransferase VII"
/label=aphVII
/note="aphVII; hygromycin B resistance"
/protein_id="AIS68763.1"
/translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
VLRVPGESTNPVLVGEPGPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
gene 3266..4264
/gene="aphVII"
/label=aphVII
3'UTR 4274..4512
/label=RBCS2 3'UTR
/note="RBCS2 3'UTR"
misc_feature 4513..6831
/label=pBluescript II KS(+)
/note="pBluescript II KS(+)"
promoter complement(4531..4549)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4570..4586)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4594..4610
/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(4618..4648)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4663..4684
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4972..5560)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5731..6591)
/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"
promoter complement(6592..6696)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6723..347)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
我们不提供该载体，载体信息仅供查询和参考。