LOCUS       sequence-359660-        5388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Tetraspanin/Membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5388)
  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 5388)
  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: CD9-mEGFP
FEATURES             Location/Qualifiers
     source          1..5388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        192..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     650..670
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      738..747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(744..746,747..749,750..1460)
                     /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(789..810)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1050..1069)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1397..1418
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2307..2428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2344..2363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2398..2417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2435..2890)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2572..2593)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2784..2803
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2917..3021
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3023..3380
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3023..3214
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3046..3066)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3231..3366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3415..4209
                     /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             3415..4209
                     /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(3469..3488)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4079..4098
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4397..4416)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4441..4488
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(4817..5388,1..17)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5306..5325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
       61 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      121 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      661 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      721 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      781 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      841 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      901 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
      961 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1021 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1081 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1141 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1201 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1261 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1321 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     1381 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1441 gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcc atgccggtca
     1501 aaggaggcac caagtgcatc aaatacctgc tgttcggatt taacttcatc ttctggcttg
     1561 ccgggattgc tgtccttgcc attggactat ggctccgatt cgactctcag accaagagca
     1621 tcttcgagca agaaactaat aataataatt ccagcttcta cacaggagtc tatattctga
     1681 tcggagccgg cgccctcatg atgctggtgg gcttcctggg ctgctgcggg gctgtgcagg
     1741 agtcccagtg catgctggga ctgttcttcg gcttcctctt ggtgatattc gccattgaaa
     1801 tagctgcggc catctgggga tattcccaca aggatgaggt gattaaggaa gtccaggagt
     1861 tttacaagga cacctacaac aagctgaaaa ccaaggatga gccccagcgg gaaacgctga
     1921 aagccatcca ctatgcgttg aactgctgtg gtttggctgg gggcgtggaa cagtttatct
     1981 cagacatctg ccccaagaag gacgtactcg aaaccttcac cgtgaagtcc tgtcctgatg
     2041 ccatcaaaga ggtcttcgac aataaattcc acatcatcgg cgcagtgggc atcggcattg
     2101 ccgtggtcat gatatttggc atgatcttca gtatgatctt gtgctgtgct atccgcagga
     2161 accgcgagat ggtctaagga tccaccggat ctagataact gatcataatc agccatacca
     2221 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2281 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2341 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2401 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     2461 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2521 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2581 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2641 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2701 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2761 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2821 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2881 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2941 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3001 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3061 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3181 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3241 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3301 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3361 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3841 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3961 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4201 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4261 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4321 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4381 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4441 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4501 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4561 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4621 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggg
//
