LOCUS       sequence-170495-        4681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for fusion of gene of interest with a monomeric reversibly 
            photoswitchable fluorescent protein, Skylan-NS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4681)
  AUTHORS   Zhang X, Zhang M, Li D, He W, Peng J, Betzig E, Xu P
  TITLE     Highly photostable, reversibly photoswitchable fluorescent protein 
            with high contrast ratio for live-cell superresolution microscopy.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Sep 13;113(37):10364-9. doi: 
            10.1073/pnas.1611038113. Epub 2016 Aug 25.
  PUBMED    27562163
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: 
            "10.1073/pnas.1611038113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-09-13- 13"; volume: "113"; issue: "37"; pages: "10364-9"
COMMENT     Sequence Label: pSkylan-NS-N1
FEATURES             Location/Qualifiers
     source          1..4681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        127..430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        431..634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     585..605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    675..731
                     /label=MCS
                     /note="multiple cloning site"
     CDS             733..1413
                     /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
                     EGGPLPFAFDILTTAFLYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDVEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    1535..1656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1572..1591)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1626..1645
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1663..2118)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1800..1821)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2012..2031
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2145..2249
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2251..2608
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2251..2442
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2274..2294)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2459..2594
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2643..3437
                     /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             2643..3437
                     /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(2697..2716)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3307..3326
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3625..3644)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3669..3716
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4045..4633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4534..4553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
       61 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      361 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      421 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      481 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      541 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      601 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      661 gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
      721 gcccgggatc caatgagtgc gattaagcca gacatgaaga tcaaactccg tatggaaggc
      781 aacgtaaacg ggcaccactt tgtgatcgac ggagatggta caggcaagcc ttttgaggga
      841 aaacagagta tggatcttga agtcaaagag ggcggacctc tgccttttgc ctttgatatc
      901 ctgaccactg cattcctgta cggcaacagg gtattcgcca aatatccaga caacatacaa
      961 gactatttta agcagtcgtt tcctaagggg tattcgtggg aacgaagctt gactttcgaa
     1021 gacgggggca tttgcaacgc cagaaacgac ataacaatgg aaggggacac tttctataat
     1081 aaagttcgat tttatggtac caactttccc gccaatggtc cagttatgca gaagaagacg
     1141 ctgaaatggg agccctccac tgagaaaatg tatgtgcgtg atggagtgct gacgggtgat
     1201 gtagagatgg ctttgttgct tgaaggaaat gcccattacc gatgtgactt cagaactact
     1261 tacaaagcta aggagaaggg tgtcaagtta ccaggcgccc actttgtgga ccactgcatt
     1321 gagattttaa gccatgacaa agattacaac aaggttaagc tgtatgagca tgctgttgct
     1381 cattctggat tgcctgacaa tgccagacga taagcggccg cgactctaga tcataatcag
     1441 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     1501 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     1561 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1621 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     1681 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1741 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1801 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1861 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1921 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1981 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2041 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2101 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2161 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2221 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2281 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2341 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2401 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2461 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2521 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2581 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     2641 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2701 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2761 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2821 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2881 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2941 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3001 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3061 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3121 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3181 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3241 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3301 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3361 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3421 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3481 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3541 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3601 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3661 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     3721 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     3781 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     3841 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     3901 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3961 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4021 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4081 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4141 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4201 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4261 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4321 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4381 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4441 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4501 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4561 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4621 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4681 t
//
