LOCUS       Exported                4716 bp ds-DNA    circular SYN 04-1-2013
DEFINITION  Vector for fusing LSSmOrange to the N-terminus of a partner protein.
ACCESSION   .
VERSION     .
KEYWORDS    pLSSmOrange-C1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4716)
  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 4716)
  AUTHORS   Verkhusha Lab
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4716
                     /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"
     CDS             607..1314
                     /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"
     misc_feature    1315..1380
                     /note="MCS"
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     misc_feature    1389..1399
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    1504..1625
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1632..2087)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2114..2218
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2220..2577
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2428..2563
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             2612..3406
                     /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    3638..3685
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      4014..4602
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"