LOCUS       sequence-370101-        5495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hetero-dimer expression vector with plasma membrane localization 
            signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5495)
  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 5495)
  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: myr-palm-mCherry2-mEGFP
FEATURES             Location/Qualifiers
     source          1..5495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /gene="bla"
                     /label=AmpR promoter
     promoter        133..490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        133..324
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(156..176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      341..476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             525..1319
                     /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             525..1319
                     /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(579..598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1189..1208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1507..1526)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1551..1598
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1927..2515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2416..2435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2690..2993
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2994..3197
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3148..3168
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3226..3235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3264..3320
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      3328..3337
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3334..4041
                     /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(3479..3497)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             join(4063..4065,4066..4068,4069..4782)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4108..4129)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4369..4388)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4716..4737
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4912..5033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4949..4968)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5003..5022
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5040..5495)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5177..5198)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5389..5408
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
      361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
      421 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
      481 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
      541 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
      601 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
      661 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
      721 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
      781 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
      841 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
      901 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
      961 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     1021 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     1081 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     1141 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     1201 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     1261 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     1321 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     1381 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     1441 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     1501 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     1561 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     1621 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     1681 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     1741 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     1801 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1861 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1921 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1981 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2041 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2101 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2161 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2221 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2281 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2341 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2401 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2461 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2521 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2581 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     2641 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2701 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2761 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2821 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2881 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2941 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     3001 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     3061 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     3121 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     3181 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgccac catgggatgc
     3241 atcaactcga agagaaagga ttcagatctc gagctcaagc ttcgaattct gcagtcgacg
     3301 gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga ggaggataac
     3361 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     3421 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     3481 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     3541 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     3601 ctgtccttcc ccgagggctt caattgggag cgcgtgatga acttcgagga cggcggcgtg
     3661 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     3721 cgcggcacca acttcccctc cgacggcccc gtaatgcagt gtcgtaccat gggctgggag
     3781 gcctccactg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     3841 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     3901 aagcccgtgc agctgcccgg cgcctacaac gtcgacatca agttggacat cctttcccac
     3961 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     4021 ggcatggacg agctgtacaa gtccggactc agatctcgag gaatggtgag caagggcgag
     4081 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     4141 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     4201 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     4261 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4321 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     4381 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     4441 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     4501 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4561 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4621 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4681 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4741 gccgccggga tcactctcgg catggacgag ctgtacaagt aaggatccac cggatctaga
     4801 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4861 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     5041 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5101 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5161 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5221 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5281 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5341 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5401 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5461 acccgccgcg cttaatgcgc cgctacaggg cgcgt
//
