pLSSmOrange-N1 载体 (V011980#)

载体基本信息
载体名称 pLSSmOrange-N1 载体抗性 Kanamycin
载体长度 4723 bp 载体类型 Fluorescent Protein Genes & Plasmids
载体来源 Shcherbakova DM, Hink MA, Joosen L, Gadella TW, Verkhusha VV. 筛选标记 Neomycin/G418(Geneticin)
拷贝数 High copy number

pLSSmOrange-N1 载体载体图谱

pLSSmOrange-N14723 bp600120018002400300036004200CMV enhancerCMV promoterMCSLSSmOrangeSV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRHSV TK poly(A) signalori

pLSSmOrange-N1 载体载体序列

LOCUS       Exported                4723 bp ds-DNA    circular SYN 04-1-2013
DEFINITION  Vector for fusing LSSmOrange to the C-terminus of a partner protein.
ACCESSION   .
VERSION     .
KEYWORDS    pLSSmOrange-N1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4723)
  AUTHORS   Shcherbakova DM, Hink MA, Joosen L, Gadella TW, Verkhusha VV.
  TITLE     An orange fluorescent protein with a large Stokes shift for 
            single-excitation multicolor FCCS and FRET imaging.
  JOURNAL   J. Am. Chem. Soc. 2012;134:7913-23.
  PUBMED    22486524
REFERENCE   2  (bases 1 to 4723)
  AUTHORS   Verkhusha Lab
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4723
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     enhancer        60..364
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    591..671
                     /note="MCS"
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     CDS             679..1389
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift"
                     /note="LSSmOrange"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    1511..1632
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1639..2094)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2121..2225
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2227..2584
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             2619..3413
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    3645..3692
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      4021..4609
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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