LOCUS       sequence-359695-        6873 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 6873)
  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 6873)
  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: mEGFP-(L)-mApple
FEATURES             Location/Qualifiers
     source          1..6873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     265..285
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(294..722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(404..425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     616..635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    768..992
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     981..998
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1084..1791)
                     /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     complement(1359..1378)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(join(1822..2532,2533..2535,2536..2538))
                     /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(1864..1885)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2213..2232
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2472..2493
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2580..2599)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2581..2599)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2644..2847)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2673..2693)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2848..3227
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3399..3418
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3493..3597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3598..3666,3667..4458)
                     /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     complement(3816..3835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4629..5217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5118..5137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5371..5388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5505..5526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5541..5546,5547..5564,5565..5571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5579..5595
                     /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)."
     primer_bind     5584..5606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5603..5619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5603..5619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5668..5789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5679..5698)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5733..5752
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5963..6757)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6074..6093)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6684..6703
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6824..6873,1..280))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6838..6873,1..100)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
       61 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
      121 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
      181 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      241 ctggggagcc tggggacttt ccacacccta actgacacac attccacaga attaattcgc
      301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
      361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
      421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
      481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
      541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
      601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
      661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
      721 gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac
      781 cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
      841 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
      901 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
      961 aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg tttaaacggg
     1021 ccctctagac tcgagcggcc gccactgtgc tggatatctg cagaattcgg accctccgcc
     1081 acccttgtac agctcgtcca tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg
     1141 ttccacgatg gtgtagtcct cgttgtggga cacgatgtcc aacttgatgt cgacgatgta
     1201 ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cggcggcgta
     1261 gtggccgccg tccttcagct tcagcctctt cttgatctcg ctcttcaggg cgccgtcctc
     1321 ggggtacatc cgctcctcgg aggcctccca gcccatggtc ttcttctgca ttacggggcc
     1381 gtcggagggg aagttggtgc cgcgcagctt caccttgtag atgaacacgc cgtcctgcag
     1441 ggaggagtcc tggttaacgt gaataatgcc gccgtcctcg aagttcatca cgcgctccca
     1501 cctgaagccc tcggggaagg acagcttgaa gtagtcgggg atgtcggctg ggtgcttaat
     1561 gtagaccttg gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg
     1621 gccacccttg gtcaccttca gcttagcggt ctgaaaggcc tcgtaggggc ggccctcgcc
     1681 ctcgccctcg atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg
     1741 catgaactcc ttgatgatgg ccatgttatt ctcctcgccc ttgctcacca tgggatccac
     1801 cggtggtgca gcggcaggcg gcttgtacag ctcgtccatg ccgagagtga tcccggcggc
     1861 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt
     1921 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     1981 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     2041 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     2101 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     2161 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     2221 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     2281 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     2341 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     2401 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     2461 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     2521 ctcgcccttg ctcaccatgg gtaccaagct taagtttaaa cgctagccag cttgggtctc
     2581 cctatagtga gtcgtattaa tttcgataag ccagtaagca gtgggttctc tagttagcca
     2641 gagagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     2701 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     2761 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     2821 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     2881 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     2941 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     3001 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     3061 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     3121 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     3181 tgaccccgta attgattact attaataact agtcaataat caatgtcaac gcgtatatct
     3241 ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt
     3301 gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc
     3361 caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc agattgtact
     3421 gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt
     3481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4141 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4741 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5401 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5461 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     5521 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     5581 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     5641 ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa
     5701 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5761 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
     5821 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     5881 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     5941 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     6001 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     6061 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     6121 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     6181 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     6241 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     6301 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     6361 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     6421 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     6481 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     6541 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     6601 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     6661 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     6721 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     6781 cttgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa aagcctaggc
     6841 ctccaaaaaa gcctcctcac tacttctgga ata
//
