pAS2hA20R562A 载体 (V009428#)

载体基本信息
载体名称 pAS2hA20R562A 载体抗性 Ampicillin
载体长度 10805 bp 载体类型 Yeast expression vector
载体来源 De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.

pAS2hA20R562A 载体载体图谱

pAS2hA20R562A10805 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105002u oriTRP1 promoterunnamed protein product; TRP1f1 oriM13 fwdCYC1 terminatorunnamed protein product; CYH2ADH1 promoterunnamed protein product; GALbdHA-epitopeunnamed protein product; hA20mADH1 terminatorM13 revlac operatorlac promoterpMB1 oriunnamed protein product; ampicillin resistanceAmpR promoter

pAS2hA20R562A 载体载体序列

LOCUS       Exported               10805 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pAS2hA20R562A, complete sequence.
ACCESSION   LT727381
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10805)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10805)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 10805)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10805
                     /organism="Yeast expression vector pAS2hA20R562A"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3771."
                     /db_xref="taxon:1945452"
     rep_origin      2..1344
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(2..982)
                     /direction=LEFT
                     /label=2mu ori
                     /note="2mu ori"
     protein_bind    complement(965..1012)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1603..1883
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1884..2558
                     /codon_start=1
                     /note="unnamed protein product; TRP1"
                     /protein_id="SJL88297.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(2661..3116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     3261..3277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3314..3561
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3855..4304)
                     /codon_start=1
                     /note="unnamed protein product; CYH2"
                     /protein_id="SJL88298.1"
                     /translation="MPSRFTKTRKHRGHVSAGKGRIGKHRKHPGGRGMAGGQHHHRINM
                     DKYHPGYFGKVGMRYFHKQQAHFWKPVLNLDKLWTLIPEDKRDQYLKSASKETAPVIDT
                     LAAGYGKILGKGRIPNVPVIVKARFVSKLAEEKIRAAGGVVELIA"
     promoter        5072..5473
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      5075..5478
                     /regulatory_class="promoter"
                     /note="ADH1"
     CDS             5501..5941
                     /codon_start=1
                     /note="unnamed protein product; GALbd"
                     /protein_id="SJL88299.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     misc_feature    5957..5983
                     /label=HA-epitope
                     /note="HA-epitope"
     CDS             5957..5983
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5999..8371
                     /codon_start=1
                     /note="unnamed protein product; hA20m"
                     /protein_id="SJL88300.1"
                     /translation="MAEQVLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIHHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRNIQATLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHATSQYMWGVQDTDLVLRKALFSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLIKMASTDTPMARSGLQYNSLEEIHIFVLCNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSHHFVPLVTLKDSGPEIRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLMVIEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSEQGRREGHAQNPMEPSVPQLSLMDVKCETPNCPFFMSVN
                     TQPLCHECSERRQKNQNKLPKLNSKPGPEGLPGMALGASRGEAYEPLAWNPEESTGGPH
                     SAPPTAPSPFLFSETTAMKCRSPGCPFTLNVQHNGFCERCHNARQLHASHAPDHTRHLD
                     PGKCQACLQDVTRTFNGICSTCFKRTTAEASSSLSTSLPPSCHQASKSDPSRLVRSPSP
                     HSCHRAGNDAPAGCLSQAARTPGDRTGTSKCRKAGCVYFGTPENKGFCTLCFIEYRENK
                     HFAAASGKVSPTASRFQNTIPCLGRECGTLGSTMFEGYCQKCFIEAQNQRFHEAKRTEE
                     QLRSSQRRDVPRTTQSTSRPKCARASCKNILACRSEELCMECQHPNQRMGPGAHRGEPA
                     PEDPPKQRCRAPACDHFGNAKCNGYCNECFQFKQMYG"
     terminator      8443..8630
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      8443..8544
                     /regulatory_class="terminator"
                     /note="ADH1T"
     primer_bind     complement(8654..8670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8678..8694
                     /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(8702..8732)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(8872..9274)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(9056..9644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9815..10675)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL88301.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(10676..10780)
                     /gene="bla"
                     /label=AmpR promoter

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