LOCUS       sequence-360031-        5380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hetero-dimer expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5380)
  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 5380)
  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: mEYFP-(L)-mApple
FEATURES             Location/Qualifiers
     source          1..5380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..54)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        226..605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        606..809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     760..780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     854..873
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        854..872
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      906..915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(912..914,915..917,918..1628)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(957..978)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1218..1237)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1565..1586
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1680..2387
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2093..2112
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2740..2759
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3044..3165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3081..3100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3135..3154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3214..3230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3214..3230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3227..3249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3238..3254
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3262..3268,3269..3286,3287..3292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3307..3328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3445..3462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3616..4204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3696..3715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4375..5166,5167..5235))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4998..5017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5236..5340)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
       61 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
      121 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
      181 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
      241 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      301 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      481 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      541 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      601 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      721 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      781 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      841 ggcttatcga aattaatacg actcactata gggagaccca agctggctag cacgatgaac
      901 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      961 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1021 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1081 cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg
     1141 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1201 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1261 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1321 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1381 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1441 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1501 ccgacaacca ctacctgagc taccagtcca aactgagcaa agaccccaac gagaagcgcg
     1561 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1621 tgtacaagcc gcctgccgct gcaccaccgg tcttaagctt ggtaccgagc tcggatccca
     1681 tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1741 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1801 gcccctacga ggcctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct
     1861 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggtctac attaagcacc
     1921 cagccgacat ccccgactac ttcaagctgt ccttccccga gggcttcagg tgggagcgcg
     1981 tgatgaactt cgaggacggc ggcattattc acgttaacca ggactcctcc ctgcaggacg
     2041 gcgtgttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     2101 tgcagaagaa gaccatgggc tgggaggcct ccgaggagcg gatgtacccc gaggacggcg
     2161 ccctgaagag cgagatcaag aagaggctga agctgaagga cggcggccac tacgccgccg
     2221 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtcg
     2281 acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa cagtacgaac
     2341 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagggt ggcggagggt
     2401 ccgaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg cttgatccgg
     2461 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2521 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2581 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2641 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     2701 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     2761 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     2821 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     2881 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     2941 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3001 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     3061 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3121 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     3181 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3241 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3301 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3361 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3421 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3481 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4021 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4201 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4321 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4381 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4441 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4501 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4561 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4621 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4681 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4741 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4801 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4861 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4921 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4981 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5041 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5101 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5161 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5221 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5341 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
//
