载体基本信息 | |||
载体名称 | LIC-pKLAC-LC1 | 载体抗性 | Ampicillin |
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载体长度 | 10540 bp | 载体类型 | Expression vector |
载体来源 | Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B. |
LIC-pKLAC-LC1 载体载体图谱
LIC-pKLAC-LC1 载体载体序列
LOCUS Exported 10540 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Expression vector LIC-pKLAC-LC1, complete sequence. ACCESSION JF327848 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10540) AUTHORS Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B. TITLE High-Throughput Protein Expression Using a Combination of Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein (IFP) Detection JOURNAL PLoS ONE 6 (4), E18900 (2011) PUBMED 21541323 REFERENCE 2 (bases 1 to 10540) AUTHORS Dortay H, Mueller-Roeber B. TITLE Direct Submission JOURNAL Submitted (14-FEB-2011) Molecular Biology, University of Potsdam, Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 14476, Germany REFERENCE 3 (bases 1 to 10540) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..10540 /organism="Expression vector LIC-pKLAC-LC1" /mol_type="other DNA" /label=Kluyveromyces lactis expression vector /note="Kluyveromyces lactis expression vector" /db_xref="taxon:1029639" misc_feature 14..16 /label=start codon /note="start codon" misc_feature 26..43 /label=6xHis tag /note="6xHis tag" CDS 26..43 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" misc_feature 53..1036 /label=infrared fluorescent protein /note="infrared fluorescent protein" CDS 53..1036 /codon_start=1 /product="bacteriophytochrome-based monomeric infrared fluorescent protein (Shu et al., 2009)" /label=IFP1.4 /translation="MARDPLPFFPPLYLGGPEITTENCEREPIHIPGSIQPHGALLTAD GHSGEVLQVSLNAATFLGQEPTVLRGQTLAALLPEQWPALQAALPPGCPDALQYRATLD WPAAGHLSLTVHRVAELLILEFEPTEAWDSIGPHALRNAMFALESAPNLRALAEVATQT VRELTGFDRVMLYKFAPDATGEMIAEARREGMQAFLGHRFPASHTPAQARALYTRHLLR LTADTRAAAVPLDPVLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGG QLWGLIVCHHQTPYVLPPDLRTTLEELGRKLSGQVQRKEAGMDELYK" misc_feature 1037..1057 /label=TEV recognition site /note="TEV recognition site" CDS 1037..1057 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" misc_feature 1076..1083 /label=PmeI recognition site /note="PmeI recognition site" misc_feature 1084..1750 /label=LIC stuffer fragment /note="LIC stuffer fragment" misc_feature 1751..1758 /label=PmeI recognition site /note="PmeI recognition site" terminator 1804..2422 /label=LAC4 terminator /note="transcription terminator for Kluyveromyces lactis LAC4" regulatory 1804..2386 /regulatory_class="terminator" /note="terminator" CDS 3146..4792 /codon_start=1 /product="acetamidase" /label=acetamidase /protein_id="AEE81072.1" /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP" promoter 5428..6084 /label=LAC4 promoter (5') /note="5' portion of the Kluyveromyces lactis LAC4 promoter" regulatory 5501..6081 /regulatory_class="promoter" /note="5' promoter region" promoter complement(6101..6119) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6133..6149) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 6157..6173 /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(6181..6211) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6226..6247 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin 6535..7123 /label=pMB1 origin /note="pMB1 origin" CDS complement(7294..8154) /codon_start=1 /product="beta-lactamase" /label=beta-lactamase /protein_id="AEE81073.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8155..8259) /gene="bla" /label=AmpR promoter rep_origin 8286..8741 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 8882..8898 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" regulatory 8908..10540 /regulatory_class="promoter" /note="3' promoter region" promoter 8912..10440 /label=LAC4 promoter (3') /note="3' portion of a modified Kluyveromyces lactis LAC4 promoter" protein_bind 10446..10462 /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)."
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