LOCUS       sequence-359657-        5117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomer expression vector with plasma membrane localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5117)
  AUTHORS   Dunsing V, Petrich A, Chiantia S
  TITLE     Multicolor fluorescence fluctuation spectroscopy in living cells via
            spectral detection.
  JOURNAL   Elife. 2021 Sep 8;10. pii: 69687. doi: 10.7554/eLife.69687.
  PUBMED    34494547
REFERENCE   2  (bases 1 to 5117)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-09-8"; volume: "10. pii: 69687. doi"; pages: " 
            10.7554/eLife.69687"
COMMENT     Sequence Label: GPI-mCherry2
FEATURES             Location/Qualifiers
     source          1..5117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      625..634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             748..1455
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(893..911)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1897..2018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1934..1953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1988..2007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2025..2480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2162..2183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2374..2393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2507..2611
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2613..2970
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2613..2804
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2636..2656)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2821..2956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3005..3799
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3005..3799
                     /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(3059..3078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3669..3688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3987..4006)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4031..4078
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4407..4995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4896..4915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc
      601 agatctcgag ctcaagctta gctagccacc atggagctct tttggagtat agtctttact
      661 gtcctcctga gtttctcctg ccgggggtca gactgggaat ctctgcagtc gacggtaccg
      721 cgggcccggg atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc
      781 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      841 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      901 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      961 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     1021 ttccccgagg gcttcaattg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     1081 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1141 accaacttcc cctccgacgg ccccgtaatg cagtgtcgta ccatgggctg ggaggcctcc
     1201 actgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1261 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1321 gtgcagctgc ccggcgccta caacgtcgac atcaagttgg acatcctttc ccacaacgag
     1381 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1441 gacgagctgt acaagtatga tccgcgccca agcagcggtc attctagaca cagatatgca
     1501 cttataccca taccattagc agtaattaca acatgtattg tgctgtatat gaatgttctt
     1561 taattgagaa gacaatttct tcatttttag gtattctgaa atgtgacaga aaaccagaca
     1621 gaaccaactc caattgattg gtaacagaag atgaagacaa cagcataact aaattatttt
     1681 aaaaactaaa aagccatctg atttctcatt tgagctttag agcacaatgg atctcgaggg
     1741 atcttccata cctaccagtt ctgcgcctgc aggtcgcggc cgcgactcta gatcataatc
     1801 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1861 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1981 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2041 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2101 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2161 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2221 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2281 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2341 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2401 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2461 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2521 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2581 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2641 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2701 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2761 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2821 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2881 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2941 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3001 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3061 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3181 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3301 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3781 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3961 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4021 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4081 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4141 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4201 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4261 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4321 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4381 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4441 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4501 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4561 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4621 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4681 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4741 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4801 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4861 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4921 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4981 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5041 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5101 gccatgcatt agttatt
//
