LOCUS       sequence-315219-        4371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses frFAST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4371)
  AUTHORS   Li C, Tebo AG, Thauvin M, Plamont MA, Volovitch M, Morin X, Vriz S, 
            Gautier A
  TITLE     A Far-Red Emitting Fluorescent Chemogenetic Reporter for In Vivo 
            Molecular Imaging.
  JOURNAL   Angew Chem Int Ed Engl. 2020 Jun 22. doi: 10.1002/anie.202006576.
  PUBMED    32568417
REFERENCE   2  (bases 1 to 4371)
  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: "Angew Chem 
            Int Ed Engl. 2020 Jun 22. doi: 10.1002/anie.202006576."
COMMENT     Sequence Label: pAG504-frFAST
FEATURES             Location/Qualifiers
     source          1..4371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      555..564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             561..935
                     /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
                     EGDITGRDPKQVIGKNLFKDVAPGTVSSGFYGKFKEGVASGNLNTMFEWMIPTSRGPTK
                     VKIHMKKALSGDSYWVFVKRV"
     CDS             942..971
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1099..1220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1136..1155)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1190..1209
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1227..1682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1364..1385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1576..1595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1709..1813
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1815..2172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1815..2006
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1838..1858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2023..2158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2207..3001
                     /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             2207..3001
                     /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(2261..2280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2871..2890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3189..3208)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3233..3280
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3609..4197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4098..4117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctgccacc atggagcatg ttgcctttgg cagtgaggac atcgagaaca
      601 ctttggccaa aatggacgac ggacaactgg atgggttggc ctttggcgca attcagctcg
      661 atggtgacgg gaatatcctg cagtacaatg ctgctgaagg agacatcaca ggcagagatc
      721 ccaaacaggt gattgggaag aacttattca aggatgttgc acctggaacg gtttcctccg
      781 ggttttacgg caaattcaag gaaggcgtag cgtcagggaa tctgaacacc atgttcgaat
      841 ggatgatacc gacaagcagg ggaccaacta aggtcaagat acacatgaag aaagcccttt
      901 ccggtgacag ctattgggtc tttgtgaaac gggtgggatc cgaacaaaag cttatttctg
      961 aagaggactt gtaataggcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1021 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1081 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1141 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1201 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     1261 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1321 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1381 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1441 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1501 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     1561 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     1621 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     1681 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1741 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1801 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     1861 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1921 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1981 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2101 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2161 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2221 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2281 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2341 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     2401 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2461 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2521 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2581 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2641 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2701 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     2761 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2821 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2881 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     2941 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3001 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3061 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3121 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3181 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3241 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3301 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3361 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3421 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     3481 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3541 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3601 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3661 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3721 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3781 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3841 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3901 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3961 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4021 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4081 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4141 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4201 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4261 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     4321 atagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
