LOCUS       sequence-260849-        5655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MAP4-iFAST in mammalian cells (microtubule labeling).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5655)
  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 5655)
  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: pAG498 MAP4-iFAST
FEATURES             Location/Qualifiers
     source          1..5655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      649..658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1930..2304
                     /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
                     VKIHMKKALSGDSYWVFVKRV"
     CDS             2311..2340
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2477..2598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2514..2533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2568..2587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2605..3060)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2742..2763)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2954..2973
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3087..3191
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3193..3550
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3193..3384
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3216..3236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3401..3536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3585..4379
                     /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             3585..4379
                     /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(3639..3658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4249..4268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4567..4586)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4611..4658
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4987..5575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5476..5495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctgc caccatggtg
      661 tcccggcaag aagaagcaaa ggctgctgta ggtgtgactg gaaatgacat cactaccccg
      721 ccaaacaagg agccaccacc aagcccagaa aagaaagcaa agcctttggc caccactcaa
      781 cctgcaaaga cttcaacatc gaaagccaaa acacagccca cttctctccc taagcaacca
      841 gctcccacca cctctggtgg gttgaataaa aaacccatga gcctcgcctc aggctcagtg
      901 ccagctgccc cacacaaacg ccctgctgct gccactgcta ctgccaggcc ttccacccta
      961 cctgccagag acgtgaagcc aaagccaatt acagaagcta aggttgccga aaagcggacc
     1021 tctccatcca agccttcatc tgccccagcc ctcaaacctg gacctaaaac caccccaacc
     1081 gtttcaaaag ccacatctcc ctcaactctt gtttccactg gaccaagtag tagaagtcca
     1141 gctacaactc tgcctaagag gccaaccagc atcaagactg aggggaaacc tgctgatgtc
     1201 aaaaggatga ctgctaagtc tgcctcagct gacttgagtc gctcaaagac cacctctgcc
     1261 agttctgtga agagaaacac cactcccact ggggcagcac ccccagcagg gatgacttcc
     1321 actcgagtca agcccatgtc tgcacctagc cgctcttctg gggctctttc tgtggacaag
     1381 aagcccactt ccactaagcc tagctcctct gctcccaggg tgagccgcct ggccacaact
     1441 gtttctgccc ctgacctgaa gagtgttcgc tccaaggtcg gctctacaga aaacatcaaa
     1501 caccagcctg gaggaggccg ggccaaggta gagaaaaaaa cagaggcagc taccacagct
     1561 gggaagcctg aacctaatgc agtcactaaa gcagccggct ccattgcgag tgcacagaaa
     1621 ccgcctgctg ggaaagtcca gatagtatcc aaaaaagtga gctacagtca tattcaatcc
     1681 aagtgtgttt ccaaggacaa tattaagcat gtccctggat gtggcaatgt tcagattcag
     1741 aacaagaaag tggacatatc caaggtctcc tccaagtgtg ggtccaaagc taatatcaag
     1801 cacaagcctg gtggaggaga tgtcaagatt gaaagtcaga agttgaactt caaggagaag
     1861 gcccaagcca aagtgggagg cggattcgcg gatccaccgg tcgccaccat gagtgtgatt
     1921 aaaccagaca tggagcatgt tgcctttggc agtgaggaca tcgagaacac tctggccaaa
     1981 atggacgacg gacaactgga tgggttggcc tttggcgcaa ttcagctcga tggtgacggg
     2041 aatatcctgc agtacaatgc tgctgaagga gacatcacag gcagagatcc caaacaggtg
     2101 attgggaaga acttcttcaa ggatgttgca cctggaacgg attctcccga gttttacggc
     2161 aaattcaagg aaggcgtagc gtcagggaat ctgaacacca tgttcgaatg gatgataccg
     2221 acaagcaggg gaccaaccaa ggtcaagata cacatgaaga aagccctttc cggtgacagc
     2281 tattgggtct ttgtgaaacg ggtgggatcc gaacaaaagc ttatttctga agaggacttg
     2341 taatagcccg ggtaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2401 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2461 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2521 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2581 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2641 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2701 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2761 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2821 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2881 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2941 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3001 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3061 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3121 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3181 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3301 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3361 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3421 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3481 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3541 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3601 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3661 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3721 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3781 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3841 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3901 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3961 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4021 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4081 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4141 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4201 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4261 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4321 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4381 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4441 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4501 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4561 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4621 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4681 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4741 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4801 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4861 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4921 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4981 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5041 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5101 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5641 tcctgcgtta tcccc
//
