LOCUS       sequence-259284-        7262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescnece reporter of beta-Actin with Skylan-S. Express in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7262)
  AUTHORS   Yi X, Son S, Ando R, Miyawaki A, Weiss S
  TITLE     Moments reconstruction and local dynamic range compression of high 
            order superresolution optical fluctuation imaging.
  JOURNAL   Biomed Opt Express. 2019 Apr 16;10(5):2430-2445. doi: 
            10.1364/BOE.10.002430. eCollection 2019 May 1.
  PUBMED    31149378
REFERENCE   2  (bases 1 to 7262)
  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: 
            "10.1364/BOE.10.002430"; journalName: "Biomed Opt Express"; date: 
            "2019-04-16- 16"; volume: "10"; issue: "5"; pages: "2430-2445"
COMMENT     Sequence Label: Skylan-S-(GGGGS)x3-beta-Actin
FEATURES             Location/Qualifiers
     source          1..7262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             921..1598
                     /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             921..1598
                     /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"
     promoter        complement(2817..2835)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2818..2835)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2855..2872)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2861..3085
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3131..3559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3218..3237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3428..3449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3568..3588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3573..3902
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3753..3888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3815..3834
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3969..4763
                     /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(4023..4042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4633..4652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4937..5058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4974..4993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5028..5047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5107..5123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5107..5123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5120..5142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5131..5147
                     /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(5155..5161,5162..5179,5180..5185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5200..5221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5338..5355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5509..6097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5589..5608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6268..7059,7060..7128))
                     /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     6891..6910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7129..7233)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta
      901 ccgagctcgg atccgccacc atgagtgcga ttaagccaga catgaagatc aaactccgta
      961 tggaaggcaa cgtaaacggg caccactttg tgatcgacgg agatggtaca ggcaagcctt
     1021 ttgagggaaa acagagtatg gatcttgaag tcaaagaggg cggacctctg ccttttgcct
     1081 ttgatatcct gaccactgca ttctcctacg gcaacagggt attcgccaaa tatccagaca
     1141 acatacaaga ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga
     1201 ctttcgaaga cgggggcatt tgcaacgcca gaaacgacat aacaatggaa ggggacactt
     1261 tctataataa agttcgattt tatggtacca actttcccgc caatggtcca gttatgcaga
     1321 agaagacgct gaaatgggag ccctccactg agaaaatgta tgtgcgtgat ggagtgctga
     1381 cgggtgatgt agagatggct ttgttgcttg aaggaaatgc ccattaccga tgtgacttca
     1441 gaactactta caaagctaag gagaagggtg tcaagttacc aggcgcccac tttgtggacc
     1501 actgcatcga gattttaagc catgacaaag attacaacaa ggttaagctg tatgagcatg
     1561 ctgttgctca ttctggattg cctgacaatg ccagacgaga attcgatggt ggaggcggtt
     1621 caggcggagg tggctctggc ggtggcggat cgatcaagct tatggatgat gatatcgccg
     1681 cgctcgtcgt cgacaacggc tccggcatgt gcaaggccgg cttcgcgggc gacgatgccc
     1741 cccgggccgt cttcccctcc atcgtggggc gccccaggca ccagggcgtg atggtgggca
     1801 tgggtcagaa ggattcctat gtgggcgacg aggcccagag caagagaggc atcctcaccc
     1861 tgaagtaccc catcgagcac ggcatcgtca ccaactggga cgacatggag aaaatctggc
     1921 accacacctt ctacaatgag ctgcgtgtgg ctcccgagga gcaccccgtg ctgctgaccg
     1981 aggcccccct gaaccccaag gccaaccgcg agaagatgac ccagatcatg tttgagacct
     2041 tcaacacccc agccatgtac gttgctatcc aggctgtgct atccctgtac gcctctggcc
     2101 gtaccactgg catcgtgatg gactccggtg acggggtcac ccacactgtg cccatctacg
     2161 aggggtatgc cctcccccat gccatcctgc gtctggacct ggctggccgg gacctgactg
     2221 actacctcat gaagatcctc accgagcgcg gctacagctt caccaccacg gccgagcggg
     2281 aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc cctggacttc gagcaagaga
     2341 tggccacggc tgcttccagc tcctccctgg agaagagcta cgagctgcct gacggccagg
     2401 tcatcaccat tggcaatgag cggttccgct gccctgaggc actcttccag ccttccttcc
     2461 tgggcatgga gtcctgtggc atccacgaaa ctaccttcaa ctccatcatg aagtgtgacg
     2521 tggacatccg caaagacctg tacgccaaca cagtgctgtc tggcggcacc accatgtacc
     2581 ctggcattgc cgacaggatg cagaaggaga tcactgccct ggcacccagc acaatgaaga
     2641 tcaagatcat tgctcctcct gagcgcaagt actccgtgtg gatcggcggc tccatcctgg
     2701 cctcgctgtc caccttccag cagatgtgga tcagcaagca ggagtatgac gagtccggcc
     2761 cctccatcgt ccaccgcaaa tgcttctagc tcgagcatgc atctggaggg ccctattcta
     2821 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     2881 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2941 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3001 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3061 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3121 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3181 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3241 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3301 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3361 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3421 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3481 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3541 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3781 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3841 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3901 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3961 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4021 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4081 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4141 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4201 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4261 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4321 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4381 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4441 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4501 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4561 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4621 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4681 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4741 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4801 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4861 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4921 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5041 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5101 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5161 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5221 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6241 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6301 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6361 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6421 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6481 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6541 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6601 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6661 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6721 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6781 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6841 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6901 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6961 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7021 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7081 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7141 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7201 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7261 cg
//
