LOCUS       Exported                4741 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  DNA damage inducible Red Fluorescent Protein for mammalian 
            expression .
ACCESSION   .
VERSION     .
KEYWORDS    p21-Reporter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4741)
  AUTHORS   Fendyur A, Varma S, Lo CT, Voldman J
  TITLE     Cell-based biosensor to report DNA damage in micro- and nanosystems.
  JOURNAL   Anal Chem. 2014 Aug 5;86(15):7598-605. doi: 10.1021/ac501412c. Epub 
            2014 Jul 7.
  PUBMED    25001406
REFERENCE   2  (bases 1 to 4741)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1021/ac501412c";
            journalName: "Anal Chem"; date: "2014-08-5- 5"; volume: "86"; issue:
            "15"; pages: "7598-605"
FEATURES             Location/Qualifiers
     source          1..4741
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             802..1497
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     polyA_signal    1620..1741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1657..1676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1711..1730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1748..2203)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1885..1906)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2097..2116
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2230..2334
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2336..2693
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2359..2379)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2544..2679
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2606..2625
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2728..3522
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2782..2801)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3392..3411
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3710..3729)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3754..3801
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4130..4718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4619..4638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"