LOCUS       sequence-353704-        5679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses pFAST at the surface of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5679)
  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 5679)
  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: pAG876 pDisplay-pFAST
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        41..420
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        421..624
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     575..595
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     669..688
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        669..687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     768..827
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             831..857
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             879..1253
                     /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             1260..1289
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1293..1439
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(1461..1478)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1467..1691
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1823..2411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1903..1922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2740..2787
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2812..2831
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3019..3813)
                     /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             complement(3019..3813)
                     /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(3130..3149)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3740..3759
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(3912..3931)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4010..4201
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4158..4178
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4261..5052,5053..5121))
                     /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     4884..4903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5122..5226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5253..5679,1..2)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5340..5359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5550..5571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttttaacaaa atattaacgc ttacaattta cgcgcgcgtt gacattgatt attgactagt
       61 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      121 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      181 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      241 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      301 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      361 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      421 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      481 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      541 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      601 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      661 atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac
      721 tagtaacggc cgccagtgtg ctggaattcg gcttggggat atccaccatg gagacagaca
      781 cactcctgct atgggtactg ctgctctggg ttccaggttc cactggtgac tatccatatg
      841 atgttccaga ttatgctggg gcccagccgg ccagatctat ggagcatgtt gcctttggca
      901 gtgaggacat cgagaacact ctggccaata tggacgacga acaactggat aggttggcct
      961 ttggcgtaat tcagctcgat ggtgacggga atatcctgct gtacaatgct gctgaagggg
     1021 acatcactgg cagagatccc aaacaggtga ttgggaagaa cttcttcaag gatgttgcac
     1081 ctggaacgga tactcccgag ttttacggca aattcaagga aggcgcagcg tcagggaatc
     1141 tgaacaccat gttcgaatgg acgataccga caagcagggg accaaccaag gtcaaggtgc
     1201 acttgaagaa agccctttcc ggtgacagat attgggtctt tgtgaaacgg gtggtcgacg
     1261 aacaaaaact catctcagaa gaggatctga atgctgtggg ccaggacacg caggaggtca
     1321 tcgtggtgcc acactccttg ccctttaagg tggtggtgat ctcagccatc ctggccctgg
     1381 tggtgctcac catcatctcc cttatcatcc tcatcatgct ttggcagaag aagccacgtt
     1441 aggcggccgc tcgagatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1501 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1561 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1621 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1681 gggctctatg gcttctgagg cggaaagaac cagtggcggt aatacggtta tccacagaat
     1741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     2581 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     2641 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     2701 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     2761 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     2821 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     2881 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     2941 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     3001 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3061 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     3121 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     3181 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     3241 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     3301 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     3361 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     3421 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     3481 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     3541 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     3601 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     3661 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     3721 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     3781 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     3841 atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     3901 cagaggccgg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     3961 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     4021 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4081 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     4141 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     4201 ggactcttcc tttttcaata aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4261 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4321 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4381 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4441 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4501 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4561 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4621 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4681 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4741 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4801 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4861 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4921 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4981 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5041 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5101 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5161 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5221 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     5281 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5341 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5401 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5461 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5521 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5581 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5641 gttaaaaaat gagctgattt aacaaaaatt taacgcgaa
//
