LOCUS       sequence-250333-        4794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FAST (also called YFAST) fused to zebrafish histone H2B in
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4794)
  AUTHORS   Plamont MA, Billon-Denis E, Maurin S, Gauron C, Pimenta FM, Specht 
            CG, Shi J, Querard J, Pan B, Rossignol J, Moncoq K, Morellet N, 
            Volovitch M, Lescop E, Chen Y, Triller A, Vriz S, Le Saux T, Jullien
            L, Gautier A
  TITLE     Small fluorescence-activating and absorption-shifting tag for 
            tunable protein imaging in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):497-502. doi: 
            10.1073/pnas.1513094113. Epub 2015 Dec 28.
  PUBMED    26711992
REFERENCE   2  (bases 1 to 4794)
  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.1513094113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-01-19- 19"; volume: "113"; issue: "3"; pages: "497-502"
COMMENT     Sequence Label: pAG109 H2B-FAST
FEATURES             Location/Qualifiers
     source          1..4794
                     /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"
     regulatory      1029..1038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1035..1409
                     /codon_start=1
                     /product="small protein tag, derived from photoactive 
                     yellow protein, that allows in vivo fluorescent labeling by
                     addition of a cell-permeant fluorogen (Plamont et al., 
                     2016)"
                     /label=Y-FAST
                     /translation="MEHVAFGSEDIENTLAKMDDGQLDGLAFGAIQLDGDGNILQYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDSPEFYGKFKEGVASGNLNTMFEWMIPTSRGPTK
                     VKVHMKKALSGDSYWVFVKRV"
     CDS             1416..1445
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1582..1703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1619..1638)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1673..1692
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1710..2165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1847..1868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2059..2078
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2192..2296
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2298..2655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2298..2489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2321..2341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2506..2641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2690..3484
                     /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             2690..3484
                     /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(2744..2763)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3354..3373
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3672..3691)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3716..3763
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4092..4680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4581..4600
                     /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 atctgaattc gccaccatgc ccgaacctgc gaagtcagcg cccgctccca
      661 aaaaaggctc taaaaaagct gtcgccaaga cccagaagaa gggggataag aaaaggcgta
      721 agaccaggaa agagagttac gccatttacg tgtacaaagt actaaaacaa gtccacccgg
      781 acactggcat ctcctcaaag gcgatgggca ttatgaactc atttgtaaac gacatcttcg
      841 agcgcatcgc cggagaagcg tcgcgcctgg cgcattacaa caagcgctcc actatcacat
      901 cccgggagat ccagacggcc gtgcgcctgc tcctgcccgg agaactggcc aaacacgctg
      961 tgtctgaggg cacaaaggcc gtgaccaagt acaccagctc caagggcgga ggctccggag
     1021 gcggatctgc caccatggag catgttgcct ttggcagtga ggacatcgag aacactctgg
     1081 ccaaaatgga cgacggacaa ctggatgggt tggcctttgg cgcaattcag ctcgatggtg
     1141 acgggaatat cctgcagtac aatgctgctg aaggagacat cacaggcaga gatcccaaac
     1201 aggtgattgg gaagaacttc ttcaaggatg ttgcacctgg aacggattct cccgagtttt
     1261 acggcaaatt caaggaaggc gtagcgtcag ggaatctgaa caccatgttc gaatggatga
     1321 taccgacaag caggggacca accaaggtca aggtgcacat gaagaaagcc ctttccggtg
     1381 acagctattg ggtctttgtg aaacgggtgg gatccgaaca aaagcttatt tctgaagagg
     1441 acttgtaata gcccgggtaa gcggccgcga ctctagatca taatcagcca taccacattt
     1501 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1561 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1621 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1681 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     1741 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1801 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1861 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1921 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1981 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2041 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2101 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2161 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2221 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2281 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2401 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2461 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2521 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2581 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2641 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2701 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2761 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2821 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     2881 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2941 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3001 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3061 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3121 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3181 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3241 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3301 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3361 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3421 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3481 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3541 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3601 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     3661 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3721 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3781 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3841 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3901 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     3961 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4021 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4081 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4141 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4201 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4261 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4321 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4381 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4441 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4501 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4561 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4621 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4681 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4741 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
