LOCUS       sequence-254634-        4681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A green photo-switchable fluorescent protein for super-resolution 
            optical fluctuation imaging (SOFI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4681)
  AUTHORS   Zhang X, Chen X, Zeng Z, Zhang M, Sun Y, Xi P, Peng J, Xu P
  TITLE     Development of a reversibly switchable fluorescent protein for 
            super-resolution optical fluctuation imaging (SOFI).
  JOURNAL   ACS Nano. 2015 Mar 24;9(3):2659-67. doi: 10.1021/nn5064387. Epub 
            2015 Feb 23.
  PUBMED    25695314
REFERENCE   2  (bases 1 to 4681)
  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"; doi: "10.1021/nn5064387";
            journalName: "ACS Nano"; date: "2015-03-24- 24"; volume: "9"; issue:
            "3"; pages: "2659-67"
COMMENT     Sequence Label: Skylan-S-N1
FEATURES             Location/Qualifiers
     source          1..4681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    609..665
                     /label=MCS
                     /note="multiple cloning site"
     CDS             667..1347
                     /codon_start=1
                     /product="truly monomeric reversibly photoswitchable 
                     fluorescent protein, suitable for nonlinear structured 
                     illumination microscopy (Zhang et al., 2016)"
                     /label=Skylan-NS
                     /note="mEos3.1 derivative that eliminates the green-to-red 
                     conversion"
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFSYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDVEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     CDS             667..1347
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.1
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFSYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDVEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    1469..1590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1506..1525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1560..1579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1597..2052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1734..1755)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1946..1965
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2079..2183
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2185..2542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2185..2376
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2208..2228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2393..2528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2577..3371
                     /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             2577..3371
                     /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(2631..2650)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3241..3260
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3559..3578)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3603..3650
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3979..4567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4468..4487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatccaatga gtgcgattaa gccagacatg aagatcaaac tccgtatgga aggcaacgta
      721 aacgggcacc actttgtgat cgacggagat ggtacaggca agccttttga gggaaaacag
      781 agtatggatc ttgaagtcaa agagggcgga cctctgcctt ttgcctttga tatcctgacc
      841 actgcattct cctacggcaa cagggtattc gccaaatatc cagacaacat acaagactat
      901 tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa gcttgacttt cgaagacggg
      961 ggcatttgca acgccagaaa cgacataaca atggaagggg acactttcta taataaagtt
     1021 cgattttatg gtaccaactt tcccgccaat ggtccagtta tgcagaagaa gacgctgaaa
     1081 tgggagccct ccactgagaa aatgtatgtg cgtgatggag tgctgacggg tgatgtagag
     1141 atggctttgt tgcttgaagg aaatgcccat taccgatgtg acttcagaac tacttacaaa
     1201 gctaaggaga agggtgtcaa gttaccaggc gcccactttg tggaccactg cattgagatt
     1261 ttaagccatg acaaagatta caacaaggtt aagctgtatg agcatgctgt tgctcattct
     1321 ggattgcctg acaatgccag acgataagcg gccgcgactc tagatcataa tcagccatac
     1381 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1441 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1501 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1561 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     1621 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1681 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1741 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1801 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1861 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1921 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1981 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2041 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2101 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2161 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     2221 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2281 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2341 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2401 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2461 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2521 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     2581 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2641 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2701 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     2761 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2821 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2881 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2941 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3001 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3061 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     3121 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3181 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3241 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3301 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3361 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3421 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3481 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3541 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     3601 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     3661 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3721 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3781 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     3841 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3901 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3961 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4021 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4081 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4141 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4201 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4261 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4321 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4381 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4441 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4501 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4561 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4621 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     4681 t
//
