pOpt_Clover_Hyg 载体 (V004221#)

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_Hyg6831 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600pBluescript II KS(+)Chlamydomonas reinhardtii heat shock protein 70A (HSP70Ap) promoterChlamydomonas reinhardtii ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit 2 promoter (RBCS2p)Chlamydomonas reinhardtii ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit 2 intron 1 (RBCS2i1)Factor Xa siteChlamydomonas reinhardtii ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit 2 intron 2 (RBCS2i2)Factor Xa siteStrep-Tag II3'UTRmultiple cloning siteHSP70ApRBCS2pRBCS2i1aphVII3'UTRpBluescript II KS(+)

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  JOURNAL   Unpublished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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