载体基本信息 | |||
载体名称 | pOpt_Clover_Hyg | 载体抗性 | Ampicillin |
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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targeted expression of nuclear transgenes in Chlamydomonas reinhardtii with a novel, modular vector toolkit JOURNAL Unpublished REFERENCE 2 (bases 1 to 6831) AUTHORS Lauersen KJ, Mussgnug JH, Kruse O. TITLE Direct Submission JOURNAL Submitted (25-JUN-2014) Algae Biotechnology and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"
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