LOCUS       Exported                8867 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSplPIGver2, complete sequence.
ACCESSION   KT831770
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8867)
  AUTHORS   Gurskaia NG, Staroverov DB, Fradkov AF, Lukianov KA.
  TITLE     [Coding region of far-red fluorescent protein katushka contains a 
            strong donor splice site]
  JOURNAL   Bioorg. Khim. 37 (3), 425-428 (2011)
  PUBMED    21899059
REFERENCE   2  (bases 1 to 8867)
  AUTHORS   Gurskaya NG, Staroverov DB, Zhang L, Fradkov AF, Markina NM, 
            Pereverzev AP, Lukyanov KA.
  TITLE     Analysis of alternative splicing of cassette exons at single-cell 
            level using two fluorescent proteins
  JOURNAL   Nucleic Acids Res. 40 (8), E57 (2012)
  PUBMED    22259036
REFERENCE   3  (bases 1 to 8867)
  AUTHORS   Gurskaya NG, Staroverov DB, Fradkov AF, Markina NM, Perverzev AP, 
            Lukyanov KA.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2015) Department of Genetics and Postgenomic 
            Technologies Laboratory of Biophotonics, M.M. Shemyakin and Yu.a. 
            Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy
            of Sciences, Ulitsa Miklukho-Maklaya, 16/10, Moscow 117997, Russian 
            Federation
REFERENCE   4  (bases 1 to 8867)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8867
                     /organism="Cloning vector pSplPIGver2"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /country="Russia: Moscow"
                     /lat_lon="55.75 N 37.60 E"
                     /db_xref="taxon:1805945"
     regulatory      1..589
                     /regulatory_class="promoter"
                     /note="CMVIE promoter"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    590..612
                     /note="multiple cloning site; enhancer; TATA box"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             join(613..1549,2728..2924,4607..5533)
                     /codon_start=1
                     /product="Katushka-PIG3-TagGFP2"
                     /label=Katushka-PIG3-TagGFP2
                     /protein_id="AMM04548.1"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHSSGLKISGNVQAGDYVLIHAGLSGVGTAAIQLTRMAGAIPLVTAGSQ
                     KKLQMAEKLGAAAGFNYKKEDFSEATLKFTKGAGVNLILDCIGGSYWEKNVNCLALDGR
                     WVLYGLMGGGDINGPLFSKLLFKRGSLITSLLRSRDNKYKQMLVNAFTEQILPHFSTEG
                     PQRLLPVLDRIYPVTEIQEAHKYMEANKNIGKIVLELPQTVDRDPPVATMSGGEELFAG
                     IVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIKFICTTGKLPVPWPTLVTTLCYGIQ
                     CFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDGKYKTRGEVKFEGDTLVNRIELKGK
                     DFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEANFKTRHNIEGGGVQLADHYQTNVP
                     LGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLESFSACCHTHGMDELYR"
     CDS             join(613..1549,4607..4734)
                     /codon_start=1
                     /product="TurboFP635-PIG3"
                     /label=TurboFP635-PIG3
                     /protein_id="AMM04549.1"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHSSGLKISGNVQAGDYVLIHAGLSGVGTAAIQLTRMAGAIPLVTAGSQ
                     KKLQMAEKLGAAAGFNYKKEDFSEATLKFTKVQANAGECFHGANSASLLHGGPPTSAAG
                     SGQNLPSDRNPGGP"
     CDS             613..1317
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     misc_feature    1318..1333
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    4790..4816
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             4817..5533
                     /codon_start=1
                     /product="TagGFP2"
                     /label=TagGFP2
                     /protein_id="AMM04547.1"
                     /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK
                     FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG
                     KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA
                     NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE
                     SFSACCHTHGMDELYR"
     polyA_signal    5655..5776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5783..6238)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6265..6369
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6371..6728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6579..6714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6763..7557
                     /codon_start=1
                     /product="neomycin-kanamycin phosphotransferase"
                     /label=neomycin-kanamycin phosphotransferase
                     /note="KanR/NeoR"
                     /protein_id="AMM04550.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    7789..7836
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8165..8753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"