LOCUS       sequence-353710-        4785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pFAST fused to zebrafish histone H2B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4785)
  AUTHORS   Benaissa H, Ounoughi K, Aujard I, Fischer E, Goiame R, Nguyen J, 
            Tebo AG, Li C, Le Saux T, Bertolin G, Tramier M, Danglot L, 
            Pietrancosta N, Morin X, Jullien L, Gautier A
  TITLE     Engineering of a fluorescent chemogenetic reporter with tunable 
            color for advanced live-cell imaging.
  JOURNAL   Nat Commun. 2021 Nov 30;12(1):6989. doi: 10.1038/s41467-021-27334-0.
  PUBMED    34848727
REFERENCE   2  (bases 1 to 4785)
  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: "Nat 
            Commun."; date: "2021-11-30"; volume: "12(1):6989. doi"; pages: " 
            10.1038/s41467-021-27334-0"
COMMENT     Sequence Label: pAG657 H2B-pFAST
FEATURES             Location/Qualifiers
     source          1..4785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        456..659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     610..630
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1120..1129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1126..1500
                     /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="MEHVAFGSEDIENTLANMDDEQLDRLAFGVIQLDGDGNILLYNAA
                     EGDITGRDPKQVIGKNFFKDVAPGTDTPEFYGKFKEGAASGNLNTMFEWTIPTSRGPTK
                     VKVHLKKALSGDRYWVFVKRV"
     CDS             1507..1536
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1664..1785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1701..1720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1755..1774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1792..2247)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1929..1950)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2141..2160
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2274..2378
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2380..2737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2380..2571
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2403..2423)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2588..2723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2772..3566
                     /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             2772..3566
                     /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(2826..2845)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3436..3455
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3754..3773)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3798..3845
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4174..4762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4663..4682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
       61 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
      121 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      541 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      601 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      661 gtttagtgaa ccgtcagatc cgctagcgct accggactca gatctgaatt cgccaccatg
      721 cccgaacctg cgaagtcagc gcccgctccc aaaaaaggct ctaaaaaagc tgtcgccaag
      781 acccagaaga agggggataa gaaaaggcgt aagaccagga aagagagtta cgccatttac
      841 gtgtacaaag tactaaaaca agtccacccg gacactggca tctcctcaaa ggcgatgggc
      901 attatgaact catttgtaaa cgacatcttc gagcgcatcg ccggagaagc gtcgcgcctg
      961 gcgcattaca acaagcgctc cactatcaca tcccgggaga tccagacggc cgtgcgcctg
     1021 ctcctgcccg gagaactggc caaacacgct gtgtctgagg gcacaaaggc cgtgaccaag
     1081 tacaccagct ccaagggcgg aggctccgga ggcggatctg ccaccatgga gcatgttgcc
     1141 tttggcagtg aggacatcga gaacactctg gccaatatgg acgacgaaca actggatagg
     1201 ttggcctttg gcgtaattca gctcgatggt gacgggaata tcctgctgta caatgctgct
     1261 gaaggggaca tcactggcag agatcccaaa caggtgattg ggaagaactt cttcaaggat
     1321 gttgcacctg gaacggatac tcccgagttt tacggcaaat tcaaggaagg cgcagcgtca
     1381 gggaatctga acaccatgtt cgaatggacg ataccgacaa gcaggggacc aaccaaggtc
     1441 aaggtgcact tgaagaaagc cctttccggt gacagatatt gggtctttgt gaaacgggtg
     1501 ggatccgaac aaaagcttat ttctgaagag gacttgtaat aggcggccgc gactctagat
     1561 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     1621 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     1681 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1741 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     1801 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1861 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     1921 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1981 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2041 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2101 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2161 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2221 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2281 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2341 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     2401 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2461 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2521 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2581 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2641 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2701 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     2761 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2821 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2881 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2941 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3001 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3061 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3121 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3181 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3241 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3301 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3361 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3421 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3481 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3541 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     3601 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     3661 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3721 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     3781 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     3841 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     3901 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     3961 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4021 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4081 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4321 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4741 aggggggcgg agcctatgga aaaacgccag caacgcggcc ttttt
//
