LOCUS       sequence-339993-        6203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single p.Arg236His(R236H) mutation was individually inserted by 
            site-directed mutagenesis into the wild-type mGFAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6203)
  AUTHORS   Letra-Vilela R, Quiteres R, Murtinheira F, Crevenna A, Hensel Z, 
            Herrera F
  TITLE     New tools for the visualization of glial fibrillary acidic protein 
            in living cells
  JOURNAL   Experimental Results (2020), Volume 1, e4, 1–8
REFERENCE   2  (bases 1 to 6203)
  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: 
            "Experimental Results (2020), Volume 1, e4, 1–8"
COMMENT     Sequence Label: mGFAP R236H
FEATURES             Location/Qualifiers
     source          1..6203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        198..501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        502..705
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     656..676
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2113..3003
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    3128..3249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3165..3184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3219..3238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3256..3711)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3393..3414)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3605..3624
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3738..3842
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3844..4201
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3844..4035
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3867..3887)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4052..4187
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4236..5030
                     /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             4236..5030
                     /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(4290..4309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4900..4919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5218..5237)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5262..5309
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(5638..6203,1..23)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6127..6146
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
       61 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      121 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
      181 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      241 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      301 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      361 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      421 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      481 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      541 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      601 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      661 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      721 cagatccgct agcgctaccg gactcagatc tcgagctcaa gcttcgaatt caggcagggc
      781 aggatggagc ggagacgcat cacctctgcg cgccgctcct atgcctccga gacggtggtc
      841 aggggcctcg gtcctagtcg acaactgggt accatgccac gcttctcctt gtctcgaatg
      901 actcctccac tccctgccag ggtggacttc tccctggccg gggcgctcaa tgctggcttc
      961 aaggagacac gggcgagcga gcgtgcagag atgatggagc tcaatgaccg ctttgctagc
     1021 tacatcgaga aggtccgctt cctggaacag caaaacaagg cgctggcagc tgaactgaac
     1081 cagcttcgag ccaaggagcc caccaaactg gctgatgtct accaggcgga gcttcgggag
     1141 ctgcggctgc ggctggacca gcttacggcc aacagtgccc ggctggaggt ggagagggac
     1201 aactttgcac aggacctcgg caccctgagg cagaagctcc aagatgaaac caacctgagg
     1261 ctggaggcag agaacaacct ggctgcgtat agacaggagg cagatgaagc caccctggct
     1321 cgtgtggatt tggagagaaa ggttgaatcg ctggaggagg agatccagtt cttaaggaag
     1381 atctatgagg aggaagttcg agaactccgg gagcagctgg cccaacagca ggtccacgtg
     1441 gagatggatg tggccaagcc agacctcaca gcggccctga gagagattca cactcaatac
     1501 gaggcagtgg ccaccagtaa catgcaagag acagaggagt ggtatcggtc taagtttgca
     1561 gacctcacag acgctgcgtc ccgcaacgca gagctgctcc gccaagccaa gcacgaagct
     1621 aacgactatc gccgccaact gcaggccttg acctgcgatc tggagtccct gcgcggcacg
     1681 aacgagtccc tagagcggca aatgcgcgaa caggaagagc gccatgcgcg ggagtcggcc
     1741 agttaccagg aggcacttgc tcggctggag gaagagggcc aaagcctcaa ggaggagatg
     1801 gcccgccacc tgcaggagta ccaggatcta ctcaacgtta agctagccct ggacatcgag
     1861 atcgccacct acaggaaatt gctggagggc gaagaaaacc gcatcaccat tcctgtacag
     1921 actttctcca acctccagat ccgagaaacc agcctggaca ccaaatccgt gtcagaaggc
     1981 cacctcaaga ggaacatcgt ggtaaagact gtggagatgc gggatggtga ggtcattaag
     2041 gactcgaagc aggagcacaa ggacgtggtg atgcagaatt ctgcagtcga cggtaccgcg
     2101 ggcccgggat ccatggcaga aatcggtact ggctttccat tcgaccccca ttatgtggaa
     2161 gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg
     2221 ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca
     2281 ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac
     2341 ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt
     2401 ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc
     2461 aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg
     2521 acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac
     2581 gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt
     2641 gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct
     2701 gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg
     2761 aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag
     2821 ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc
     2881 aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa
     2941 gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtcgacgct cgagatttcc
     3001 ggctaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     3061 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3121 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3181 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3241 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3301 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3361 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3421 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3481 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3541 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3601 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3661 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3721 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3901 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4021 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4081 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4141 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4201 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4261 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4321 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4381 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4441 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4501 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4561 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4621 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4681 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4741 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4801 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4861 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4921 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4981 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5041 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5101 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5161 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5221 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5281 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5341 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5401 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5461 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5521 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5581 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6181 gcgtcgattt ttgtgatgct cgt
//
