LOCUS       sequence-310493-        4371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses greenFAST in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4371)
  AUTHORS   Tebo AG, Moeyaert B, Thauvin M, Carlon-Andres I, Boken D, Volovitch 
            M, Padilla-Parra S, Dedecker P, Vriz S, Gautier A
  TITLE     Orthogonal fluorescent chemogenetic reporters for multicolor 
            imaging.
  JOURNAL   Nat Chem Biol. 2020 Aug 10. pii: 10.1038/s41589-020-0611-0. doi: 
            10.1038/s41589-020-0611-0.
  PUBMED    32778846
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: "Nat Chem 
            Biol. 2020 Aug 10. pii: 10.1038/s41589-020-0611-0. doi: 
            10.1038/s41589-020-0611-0."
COMMENT     Sequence Label: pAG364-greenFAST
FEATURES             Location/Qualifiers
     source          1..4371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             621..995
                     /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="MEHVAFGSEDIENTLAKMDDEQLDGLAFGAIQLDGDGNILQYNAA
                     EGDITGRDPKQVIGKNFFKDVATGTDSPEFYRKFKEGVASGNLNTMFEWMIPTSRGPTK
                     VKVHMKKALSGDSYWVFVKRV"
     CDS             1002..1031
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    1159..1280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1196..1215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1250..1269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1287..1742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1424..1445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1636..1655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1769..1873
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1875..2232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1875..2066
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1898..1918)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2083..2218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2267..3061
                     /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             2267..3061
                     /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(2321..2340)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2931..2950
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3249..3268)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3293..3340
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3669..4257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4158..4177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctgccacc atggagcatg ttgcctttgg cagtgaggac atcgagaaca
      661 ctctggccaa aatggacgac gaacaactgg atgggttggc ctttggcgca attcagctcg
      721 atggtgacgg gaatatcctg cagtacaatg ctgctgaagg agacatcaca ggcagagatc
      781 ccaaacaggt gattgggaag aacttcttca aggatgttgc aactggaacg gattctcccg
      841 agttttaccg caaattcaag gaaggcgtag cgtcagggaa tctgaacacc atgttcgaat
      901 ggatgatacc gacaagcagg ggaccaacca aggtcaaggt gcacatgaag aaagcccttt
      961 ccggtgacag ctattgggtc tttgtgaaac gggtgggatc cgaacaaaag cttatttctg
     1021 aagaggactt gtaataggcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1081 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1141 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1201 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1261 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     1321 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1381 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1441 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1501 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1561 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     1621 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     1681 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     1741 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1801 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1861 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     1921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1981 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2041 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2101 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2161 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2221 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2281 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2341 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2401 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     2461 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     2521 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     2581 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     2641 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     2701 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     2761 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     2821 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     2881 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     2941 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3001 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3061 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3121 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3181 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3241 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3301 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3361 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3421 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3481 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     3541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3601 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3661 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3721 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3781 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3841 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3901 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3961 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4021 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4081 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4141 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4201 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4261 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4321 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
