LOCUS       Exported                4780 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Plasmid expresses endoplasmic reticulum targeted mRFP in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    ER-mRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4780)
  AUTHORS   Snapp EL, Sharma A, Lippincott-Schwartz J, Hegde RS
  TITLE     Monitoring chaperone engagement of substrates in the endoplasmic 
            reticulum of live cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Apr 25;103(17):6536-41. Epub 2006 Apr
            14.
  PUBMED    16617114
REFERENCE   2  (bases 1 to 4780)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4780
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     5'UTR           536..578
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             587..676
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVS"
     CDS             689..1363
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(819..837)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1087..1106
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1281..1304
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1508..1629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1545..1564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1599..1618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1636..2091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1773..1794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1985..2004
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2118..2222
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2224..2581
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2247..2267)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2432..2567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2494..2513
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2616..3410
                     /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(2670..2689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3280..3299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3598..3617)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3642..3689
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcttgt
      541 tctttttgca gaagctcaga ataaacgctc aactttggca gataccatgg acagcaaagg
      601 ttcgtcgcag aaagggtccc gcctgctcct gctgctggtg gtgtcaaatc tactcttgtg
      661 ccagggtgtg gtctcctgca gtcgacttat ggcctcctcc gaggacgtca tcaaggagtt
      721 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      781 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      841 cggccccctg cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc
      901 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
      961 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1021 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1081 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta
     1141 ccccgaggac ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg
     1201 ccactacgac gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg
     1261 cgcctacaag accgacatca agctggacat cacctcccac aacgaggact acaccatcgt
     1321 ggaacagtac gagcgcgccg agggccgcca ctccaccggc gccaaagatg agctctaagg
     1381 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     1441 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1501 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1621 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     1681 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     1741 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     1801 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     1861 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     1921 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1981 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2041 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2101 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2161 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2221 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2281 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2341 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2401 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2461 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2521 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2581 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2641 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2701 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2761 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     2821 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2881 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2941 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3001 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3061 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3121 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3181 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3241 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3301 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3361 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3421 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3481 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3541 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     3601 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     3661 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3721 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3781 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3841 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     3901 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3961 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4681 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     4741 ttacggggtc attagttcat agcccatata tggagttccg
//
