LOCUS       sequence-315223-        5646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MAP4-frFAST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5646)
  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 5646)
  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: pAG502-MAP4-frFAST
FEATURES             Location/Qualifiers
     source          1..5646
                     /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             1836..2210
                     /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             2217..2246
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2374..2495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2411..2430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2465..2484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2502..2957)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2639..2660)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2851..2870
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2984..3088
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3090..3447
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3090..3281
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3113..3133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3298..3433
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3482..4276
                     /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             3482..4276
                     /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(3536..3555)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4146..4165
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4464..4483)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4508..4555
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4884..5472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5373..5392
                     /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 atggtgtccc ggcaagaaga agcaaaggct gctgtaggtg
      601 tgactggaaa tgacatcact accccgccaa acaaggagcc accaccaagc ccagaaaaga
      661 aagcaaagcc tttggccacc actcaacctg caaagacttc aacatcgaaa gccaaaacac
      721 agcccacttc tctccctaag caaccagctc ccaccacctc tggtgggttg aataaaaaac
      781 ccatgagcct cgcctcaggc tcagtgccag ctgccccaca caaacgccct gctgctgcca
      841 ctgctactgc caggccttcc accctacctg ccagagacgt gaagccaaag ccaattacag
      901 aagctaaggt tgccgaaaag cggacctctc catccaagcc ttcatctgcc ccagccctca
      961 aacctggacc taaaaccacc ccaaccgttt caaaagccac atctccctca actcttgttt
     1021 ccactggacc aagtagtaga agtccagcta caactctgcc taagaggcca accagcatca
     1081 agactgaggg gaaacctgct gatgtcaaaa ggatgactgc taagtctgcc tcagctgact
     1141 tgagtcgctc aaagaccacc tctgccagtt ctgtgaagag aaacaccact cccactgggg
     1201 cagcaccccc agcagggatg acttccactc gagtcaagcc catgtctgca cctagccgct
     1261 cttctggggc tctttctgtg gacaagaagc ccacttccac taagcctagc tcctctgctc
     1321 ccagggtgag ccgcctggcc acaactgttt ctgcccctga cctgaagagt gttcgctcca
     1381 aggtcggctc tacagaaaac atcaaacacc agcctggagg aggccgggcc aaggtagaga
     1441 aaaaaacaga ggcagctacc acagctggga agcctgaacc taatgcagtc actaaagcag
     1501 ccggctccat tgcgagtgca cagaaaccgc ctgctgggaa agtccagata gtatccaaaa
     1561 aagtgagcta cagtcatatt caatccaagt gtgtttccaa ggacaatatt aagcatgtcc
     1621 ctggatgtgg caatgttcag attcagaaca agaaagtgga catatccaag gtctcctcca
     1681 agtgtgggtc caaagctaat atcaagcaca agcctggtgg aggagatgtc aagattgaaa
     1741 gtcagaagtt gaacttcaag gagaaggccc aagccaaagt gggaggcgga ttcgcggatc
     1801 caccggtcgc caccatgagt gtgattaaac cagacatgga gcatgttgcc tttggcagtg
     1861 aggacatcga gaacactttg gccaaaatgg acgacggaca actggatggg ttggcctttg
     1921 gcgcaattca gctcgatggt gacgggaata tcctgcagta caatgctgct gaaggagaca
     1981 tcacaggcag agatcccaaa caggtgattg ggaagaactt attcaaggat gttgcacctg
     2041 gaacggtttc ctccgggttt tacggcaaat tcaaggaagg cgtagcgtca gggaatctga
     2101 acaccatgtt cgaatggatg ataccgacaa gcaggggacc aactaaggtc aagatacaca
     2161 tgaagaaagc cctttccggt gacagctatt gggtctttgt gaaacgggtg ggatccgaac
     2221 aaaagcttat ttctgaagag gacttgtaat aggcggccgc gactctagat cataatcagc
     2281 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2341 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2401 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2461 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2521 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2581 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2641 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2701 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2761 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2821 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2881 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2941 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3001 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3061 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3121 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3241 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3301 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3361 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3421 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3481 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3541 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3601 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3661 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3721 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3781 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3841 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3901 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3961 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4021 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4081 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4141 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4201 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4261 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4321 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4381 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4441 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4501 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4561 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4621 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4681 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4741 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4801 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4861 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4921 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4981 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5041 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5101 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5161 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5221 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5281 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5341 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5401 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5461 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5521 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5581 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5641 gttccg
//
